Dnai for the modulation of genes

ABSTRACT

The present invention relates to methods and compositions for the inhibition of gene expression. In particular, the present invention provides oligonucleotide-based therapeutics for the inhibition genes implicated in many diseases.

PRIORITY CLAIM

This application claims priority to U.S. Provisional Patent Application No. 61/794,778 filed on Mar. 15, 2013. The entire contents of the aforementioned application are incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to methods and compositions for the inhibition of gene expression. In particular, the present invention provides oligonucleotide-based therapeutics for the inhibition or interference of genes involved and implicated in diseases and cell systems.

SEQUENCE LISTING

This application incorporates by reference in its entirety the Sequence Listing entitled “DNAi13728_ST25.txt” (2.90 MB), which was created Mar. 14, 2014 and filed electronically herewith.

BACKGROUND OF THE INVENTION

The expression of gene products in cancer, e.g. oncogenes has become the central concept in understanding cancer biology and may provide valuable targets for therapeutic drugs. All oncogenes and their products operate inside the cell making protein-based drugs ineffective since their specificity involves ligand-receptor recognition.

Aside from oncogenes, proteins implicated in tumor suppression, genesis, progression, growth, proliferation, migration, cell cycle, cell signaling, metastases, invasion, transformation, differentiation, tolerance, vascular leakage, epithelial mesenchymal transition (EMT), aggregation, angiogenesis, adhesion, development of resistance, addiction to oncogenes and non-oncogenes (cytokines, chemokines, growth factors), alteration of immune surveillance or immune response, alteration of tumor stroma/local environment, endothelial activation, extracellular matrix remodeling, hypoxia and inflammation, immune activation or immune suppression, and survival and/or prevention of cell death by apoptosis, necrosis, or autophagy may be useful targets. Proteins implicated may be increased, decreased, or altered to have an impact on diseases and/or cell systems.

Similarly numerous protein products implicated (overexpressed, mutated, or suppressed) in non-cancerous diseases involving bacterial, cardiovascular (heart failure, atherosclerosis, dylipidemia, etc.), vascular, metabolic, diabetic, dental, oral, dermatological, endocrinology, fungal, gastroenterological, bowel (e.g. Crohn's, Ulcerative Colitis, or inflammatory bowel disease, etc.), genetic, hematological, hepatic, immunology, infections and/or infectious disease, inflammation (e.g. arthritis, etc.), musculosketal, nephrology, neurology (e.g. Alzheimer's, Parkinson's, Huntington's, Multiple Sclerosis, etc.), nutrition and/or weight loss, obstetrics/gynecology, ophthalmology, orthopedics, otolaryngology, pediatric/neonatology, podiatry, pulmonary/respiratory disease, rheumatology, sleep disorders, trauma, urology, stem cells, and viral (e.g. HCV, HIV, HBV, Herpes, etc.) may be useful targets.

Antisense oligonucleotides are under investigation as therapeutic compounds for specifically targeting oncogenes (Wickstrom, E. (ed). Prospects for antisense nucleic acid therapy of cancer and Aids. New York: Wiley-Liss, Inc. 1991; Murray, J. A. H. (ed). Antisense RNA and DNA New York: Wiley-Liss, Inc. 1992). Antisense drugs are modified synthetic oligonucleotides that work by interfering with ribosomal translation of the target mRNA. The antisense drugs developed thus far destroy the targeted mRNA by binding to it and triggering ribonuclease H (RNase H) degradation of mRNA. Oligonucleotides have a half-life of about 20 minutes and they are therefore rapidly degraded in most cells (Fisher, T. L. et al., Nucleic Acids Res. 21:3857-3865 (1993)). To increase the stability of oligonucleotides, they are often chemically modified, e.g., they are protected by a sulfur replacing one of the phosphate oxygens in the backbone (phosphorothioate) (Milligan, J. F. et al., J. Med. Chem. 36:1923-1937 (1993); Wagner, R. W. et al., Science 260:1510-1513 (1993)). However, this modification can only slow the degradation of antisense and therefore large dosages of antisense drug are required to be effective.

Despite the optimism surrounding the use of antisense therapies, there are a number of serious problems with the use of antisense drugs such as difficulty in getting a sufficient amount of antisense into the cell, non-sequence-specific effects, toxicity due to the large amount of sulfur containing phosphothioates oligonucleotides and their inability to enter their target cells, and their high cost due to continuous delivery of large doses. An additional problem with antisense drugs has been their nonspecific activities. Improvements to these first generation RNA targeted nucleic acid therapeutics utilize chemical modification to prevent degradation and utilize other modifications (e.g. 2′OMe modifications, CEt, locked nucleic acids (LNA), unlocked nucleic acids, inverted bases, conformationally-restricted nucleic acids (CRN)) to enable therapeutic windows of activity to be improved.

Other nucleic acid-based approaches beyond antisense also target RNA and its translational machinery rather than genomic DNA. These include double-stranded siRNA to block the translation of abberant proteins, RNA modulation to correct gene defects by exon skipping, and double or single-stranded microRNAs that function to regulate the expression of several gene pathways through the action of miRs and antimiRs, which replace absent sequences or antagonize sequences, respectively.

There is a need for additional non-protein based cancer therapeutics that target genes implicated in diseases. Therapeutics that are effective in low doses and that are non-toxic to the subject are particularly needed.

SUMMARY OF THE INVENTION

The present invention relates to methods and compositions for the interference (inhibition, enhancement or alteration) of gene transcription or gene expression. In particular, the present invention provides oligonucleotide-based therapeutics for the modulation of disease causing genes.

An oligonucleotide that hybridizes to a non-coding region of a target gene, wherein the oligonucleotide comprises: a length of 20-34 bases; at least one CG pairs; at least 40% C and G content; no more than five consecutive bases of the same nucleotide; and may form at least one secondary structure. This oligonucleotide can also comprise a C and G content of at least 30% and in some embodiments the oligonucleotide comprises a C and G content of from about 50 to 80%. In some embodiments the oligonucleotide comprises at least two CG pairs. In some embodiments the oligonucleotide is complementary of said non-coding region of the target gene. In some embodiments the oligonucleotide is unique to the nucleotide sequence of the non-coding region. In some embodiments the nucleotide sequence of the non-coding region is not duplicated in a genome comprising the target gene. In some embodiments the nucleotide sequence of the non-coding region comprises 60% or greater homology to other nucleotide sequences in a genome with another gene. In some other embodiments the oligonucleotide is complementary to a non-coding region of another gene that influences that target gene. In yet other embodiments the oligonucleotide is complementary to a non-coding region of another gene that influences that target gene due to a chromosomal rearrangement. In yet other embodiments the oligonucleotide is complementary to a region upstream of the transcription start site.

In some embodiments, the present invention provides a composition comprising one or more distinct oligonucleotides that hybridizes under physiological conditions to regions upstream of the transcription start site of a disease causing gene.

In some embodiments, the region or regions upstream of the start site are located in regions on, surrounding or near transcription factor binding sites. In other embodiments, the regions are located on, surrounding or near various classes of regulatory elements (promoters, proximal promoters, distal enhancers, activators/co-activators, suppressors) that serve as cis-regulatory elements involved in gene transcription.

In some embodiments, the present invention provides compositions that are complementary to residues within CG regions. In some other embodiments, the present invention provides compositions that are complementary to residues within CpG islands. In yet other embodiments, the present invention resides in areas within nuclease hypersensitive areas.

In some embodiments, the present invention provides a composition comprising a first oligonucleotide that hybridizes under physiological conditions to the regulatory region of the target sequences. In some embodiments, at least one of the cytosine bases in the first oligonucleotide is 5-methylcytosine. In some of the embodiments, wherein at least one or all the cytosine bases in said CG pair is 5-methylcytosine. In some embodiments, all of the cytosine bases in the first oligonucleotide are 5-methylcytosine. In yet other embodiments, some of the bases in the first oligonucleotide are modified to prevent nuclease degradation during cell culture experiments. In some preferred embodiments, the hybridization of the first oligonucleotide to the promoter region of a gene modulates expression of the target gene. In some embodiments, the target gene is on a chromosome of a cell, and the hybridization of the first oligonucleotide to the regulatory region of the gene modulates cell signaling pathways of the cell. In some embodiments, the composition further comprises a second oligonucleotide. In some embodiments, at least one (e.g. all) of the cytosines in the second oligonucleotide are 5-methylcytosine.

In yet other embodiments, the present invention provides a method, comprising: providing an oligonucleotide; and a cell capable of transcription, and a cell capable of gene expression, and comprising a gene capable of being transcribed, and comprising a gene capable of being expressed; and introducing the oligonucleotide to the cell. In some embodiments, the introducing results in the modulation of the gene transcription. In some embodiments, the introducing results in the modulation of expression of the gene. In other embodiments, the introducing results in the modulation of proliferation of the cell. In yet other embodiments, the introducing results in the modulation of the cell phenotype. In certain embodiments, the introducing results in alteration of expression of other genes related to the target gene. In certain other embodiments, the introducing results in modulation of cell signaling pathways related to the target gene transcription. In yet other embodiments, the introducing results in an interference with the expression of other genes involved in transcription. In some embodiments, the cell is a cancer cell. In other embodiments, the cell is a prokaryote. In some other embodiments, the cell is a eukaryote. In some other embodiments the cell is in a host plant. In other embodiments, the cell is in a host animal (e.g., a non-human mammal or a human). In some embodiments, the oligonucleotide is introduced to the host animal at a dosage of between 0.1 mg to 10 g, and preferably at a dosage of between 00.1 mg to 100 mg per kg of body weight or 1 to 500 mg per meter squared body surface area. In some embodiments, the oligonucleotide is introduced to the host animal one or more times per day. In other embodiments, the oligonucleotide is introduced to the host animal continuously. In still further embodiments, the cell is in cell culture. In some embodiments, the method further comprises the step of introducing a test compound to the cell. In some embodiments, the test compound is a known chemotherapy or therapeutic agent. In some embodiments, the cancer is pancreatic cancer, colon/gastric cancer, breast cancer, renal/bladder cancer, lung cancer, leukemia, prostate, lymphoma, ovarian, thyroid cancer, sarcoma, or melanoma. In some embodiments, the non cancer disease involves bacterial, cardiovascular (heart failure, atherosclerosis, dylipidemia, etc.), vascular, metabolic, diabetic, dental, oral, dermatological, endocrinology, fungal, gastroenterological, bowel (e.g. Crohn's, Ulcerative Colitis, or inflammatory bowel disease, etc.), genetic, hematological, hepatic, immunology, infections and/or infectious disease, inflammation (e.g. arthritis, etc.), musculosketal, nephrology, neurology (e.g. Alzheimer's, Parkinson's, Huntington's, Multiple Sclerosis, etc.), nutrition and/or weight loss, obstetrics/gynecology, ophthalmology, orthopedics, otolaryngology, pediatric/neonatology, podiatry, pulmonary/respiratory disease, rheumatology, sleep disorders, trauma, urology, or viral (e.g. HCV, HIV, HBV, Herpes, etc.) disease.

In some embodiments, the method further provides a drug delivery system. In some embodiments, the drug delivery system comprises a nanoparticle, nanocrystal or complex, (e.g., a liposome comprising a neutral lipid or a lipid like compound or particles comprising polymer or polymer-like compound). In some embodiments, the drug delivery system comprises a cell targeting component (e.g., a ligand or ligand like molecule for a cell surface receptor or a nuclear receptor). In yet other embodiments, the drug delivery system comprises a device to administer the test compound(s). In certain embodiments, the drug delivery system is for use in vivo, and the oligonucleotide and the liposome, nanoparticle, nanocrystal or delivery system are present in the ratio of from 1:1 to 1:1000 (weight per weight).

The present invention further provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the coding strand of a gene under conditions such that expression of that gene is inhibited, enhanced or altered (i.e. modulated)

The present invention further provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the coding strand of a gene under conditions such that transcription of that gene is inhibited, enhanced or altered (i.e. modulated)

The present invention further provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the coding strand of a gene under conditions such that cell signaling pathways related to that gene is inhibited, enhanced or altered (i.e. modulated).

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the promoter region of a gene on a chromosome of a cell under conditions such that the cell phenotype is altered.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the promoter region of a gene on a chromosome of a cell under conditions such that proliferation of the cell is reduced.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the CG regions of a gene on a chromosome of a cell under conditions such that cell signaling pathways are modulated.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to CpG islands of a gene on a chromosome of a cell under conditions such that cell signaling pathways are modulated.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the CG regions of a gene on a chromosome of a cell under conditions such that genes related to transcription of that gene are modulated.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the CpG islands of a gene on a chromosome of a cell under conditions such that genes related to gene expression of that gene are modulated.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the CG regions of a gene on a chromosome of a cell under conditions such that genes related to cell phenotype are modulated.

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the CpG islands of a gene on a chromosome of a cell under conditions such that genes related to cell phenotype are modulated.

The present invention additionally provides a method of inhibiting the expression of a gene in a subject (e.g., for the treatment of cancer or other hyperproliferative/overexpressive gene disorders) comprising providing an oligonucleotide that hybridizes under physiological conditions to the coding strand of a gene involved in cancer or a hyperproliferative/overexpressive gene disorder expressed in the biological sample, the oligonucleotide comprising at least on CG dinucleotide pair; and administering the oligonucleotide to the subject under conditions such that transcription or expression of the gene is inhibited, enhanced or altered (i.e. modulated). In some embodiments, the subject is a human.

In some embodiments, the method further provides a drug delivery system. In some embodiments, the drug delivery system comprises a liposome (e.g., a liposome comprising a neutral lipid or a lipid like compound or particles comprising polymer or polymer-like compound). In some embodiments, the drug delivery system comprises a cell targeting component (e.g., a ligand or ligand like molecule for a cell surface receptor or a nuclear receptor). In certain embodiments, the drug delivery system is for use in vivo, and the oligonucleotide and the liposome, nanoparticle, nanocrystal or delivery system are present in the ratio of from 1:1 to 1:1000 (weight per weight).

The present invention additionally provides a composition comprising an oligonucleotide that hybridizes under physiological conditions to the promoter region of a gene located on a chromosome of a cell under conditions such that transcription, phenotype or cell signaling pathways related to the target gene are modulated.

In certain embodiments, the present invention provides a kit comprising an oligonucleotide that hybridizes under physiological conditions to the promoter region of a gene, the oligonucleotide comprising at least one CG dinucleotide pair, wherein at least one of the cytosine bases in the CG dinucleotide pair comprises 5-methylcytosine; and instructions for using the kit for reducing proliferation of a cell comprising a gene on a chromosome of the cell or inhibiting gene expression. In some embodiments, the composition in the kit is used for treating cancer in a subject and the instructions comprise instructions for using the kit to treat cancer in the subject. In some embodiments, the instructions are instructions required by the U.S. Food and Drug Agency for labeling of pharmaceuticals.

The present invention also provides a method, comprising: providing a biological sample from a subject diagnosed with a cancer; and reagents for detecting the present or absence of expression of a oncogene in the sample; and detecting the presence or absence of expression of an oncogene in the sample; administering an oligonucleotide that hybridizes under physiological conditions to the promoter region of an oncogene expressed in the biological sample, the oligonucleotide comprising at least one CG dinucleotide pair.

The present invention additionally provides a method of inhibiting the expression of a gene in a subject (e.g., for the treatment of cancer or other hyperproliferative disorders) comprising providing an oligonucleotide that hybridizes under physiological conditions to the promoter region of a gene involved in cancer or a hyperproliferative disorder expressed in the biological sample, the oligonucleotide comprising at least one CG dinucleotide pair; and administering the oligonucleotide to the subject under conditions such that expression of the gene is inhibited. In some embodiments, the subject is a human.

The present invention additionally provides a method of modulating the transcription of a gene in a subject (e.g., for the treatment of disease) comprising an oligonucleotide that hybridizes under physiological conditions to the non-coding region of a gene involved in disease expressed in the biological sample, the oligonucleotide comprising at least one CG dinucleotide pair; and administering the oligonucleotide to the subject under conditions such that expression of the gene is inhibited. In some embodiments, the subject is a human.

In yet further embodiments, the present invention provides a method of screening compounds providing a cell comprising a suspected gene; and an oligonucleotide that hybridizes to the promoter region of the gene; and administering the oligonucleotide to the cell; and determining if the phenotype of the cell is modulated in the presence of the oligonucleotide relative to the absence of the oligonucleotide. In some embodiments, the cell is in culture (e.g., a prokaryote or eukaryote cell line). In other embodiments, the cell is in a host animal (e.g., a non-human mammal). In some embodiments, the method is a high-throughput screening method.

In other embodiments, the present invention relates to methods and compositions for cancer therapy. In particular, the present invention provides nanoparticle, nanocrystal, liposome, or complex based cancer or non-cancer therapeutics.

Accordingly, in some embodiments, the present invention provides a pharmaceutical composition comprising (e.g., consisting of) a cationic, neutral, or anionic lipids, polymers or delivery agents in a complex or mixture with an oligonucleotide. In some preferred embodiments, the liposome is cationic, neutral, anionic or amphoteric (e.g. SMARTICLES) in charge. In some preferred embodiments, the complex is a mixture of lipids, lipid-like, polymer or polymer-like delivery agents and a cation (e.g. lipids and calcium to form cochleates) or a mixture of lipids lipids, lipid-like, polymer or polymer-like delivery agents and an anion.

In some embodiments, the present invention provides a kit, comprising an oligonucleotide (e.g., an oligonuculeotide that hybridizes to the CG regions, CpG islands or promoter region of an onocogene) and a first pharmaceutical composition comprising (e.g., consisting of) a cationic, neutral, or anionic liposome comprises an optional second pharmaceutical composition, wherein the second pharmaceutical composition comprises a known chemotherapy agent (e.g., TAXOTERE, TAXOL, or VINCRISTINE, etc.), or chemotherapy cocktail, and wherein the known chemotherapy agent is formulated separately from the first pharmaceutical composition. In some embodiments, the chemotherapy agent is present at less than one half the standard dose, more preferably less than one third, even more preferably less than one fourth and still more preferable less than one tenth, and yet more preferably less than one hundredth the standard dose.

In some embodiments, the present invention provides a kit, comprising an oligonucleotide (e.g., an oligonuculeotide that hybridizes to the CG regions, CG islands, or promoter region of an onocogene) and a first pharmaceutical composition comprising (e.g., consisting of) a cationic, neutral, or anionic liposome comprises an optional second pharmaceutical composition, wherein the second pharmaceutical composition comprises a known agent (e.g., an antibiotic, an antiviral, an anti-inflammatory, etc.), or treatment cocktail, and wherein the known agent is formulated separately from the first pharmaceutical composition. In some embodiments, the agent is present at less than one half the standard dose, more preferably less than one third, even more preferably less than one fourth and still more preferable less than one tenth, and yet more preferably less than one hundredth the standard dose.

In yet other embodiments, the present invention provides a method, comprising providing a pharmaceutical composition consisting of a cationic, neutral, or anionic liposome and an oligonucleotide (e.g., an oligonuculeotide that hybridizes to the promoter region of an onocogene); and exposing the pharmaceutical composition to a cancer cell. In some preferred embodiments, the liposome is a cardiolipin based cationic liposome (e.g., NEOPHECTIN). In some preferred embodiments, the charge ration of NEOPHECTIN to oligonucleotide is 6:1. In other embodiments, the liposome comprises N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate (DOTAP). In some embodiments, the cancer cell is a prostate cancer cell, an ovarian cancer cell, a breast cancer cell, a leukemia cell, or lymphoma cell. In some embodiments, the cell is in a host animal (e.g., a human). In some embodiments, the pharmaceutical composition is introduced to the host animal one or more times per day (e.g., continuously). In some embodiments, the method further comprises the step of administering a known chemotherapeutic agent to the subject (e.g., TAXOTERE, TAXOL, or VINCRISTINE), wherein the known chemotherapeutic agent is formulated separately from the cationic, neutral or anionic liposome. In preferred embodiments, the known chemotherapeutic agent is administered separately from the pharmaceutical composition. In some embodiments, the chemotherapy agent is present at less than one half the standard dose, more preferably less than one third, even more preferably less than one forth and still more preferable less than one tenth, and yet more preferably less than one hundredth the standard dose.

DESCRIPTION OF THE FIGURES

FIG. 1 demonstrates a dose-dependent response for representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 2 demonstrates a dose-dependent response for representative olionucleotides in A549 (human lung cell line).

FIG. 3 demonstrates a dose-dependent response for representative olionucleotides in DU145 (human prostate cell line).

FIG. 4 demonstrates a dose-dependent response for representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 5 depicts the structure of the olionucleotide SU1.

FIG. 6 depicts the structure of the olionucleotide SU2.

FIG. 7 depicts the structure of the olionucleotide SU3.

FIG. 8 depicts the structure of the olionucleotide SU1_(—)02.

FIG. 9 depicts the structure of the olionucleotide SU1_(—)03.

FIG. 10 demonstrates target inhibition of representative olionucleotides in DU145 (human prostate cell line).

FIG. 11 demonstrates target inhibition of representative olionucleotides in HCT-116 (human colorectal carcinoma).

FIG. 12 depicts the structure of the olionucleotide BE1.

FIG. 13 depicts the structure of the olionucleotide BE2.

FIG. 14 demonstrates target inhibition of representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 15 demonstrates target inhibition of representative olionucleotides in DU145 (human prostate cell line).

FIG. 16 depicts the structure of the olionucleotide ST1.

FIG. 17 depicts the structure of the olionucleotide ST2.

FIG. 18 demonstrates target inhibition of representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 19 demonstrates target inhibition of representative olionucleotides in DU145 (human prostate cell line).

FIG. 20 depicts the structure of the olionucleotide HI1.

FIG. 21 depicts the structure of the olionucleotide HI2.

FIG. 22 demonstrates target inhibition of representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 23 demonstrates target inhibition of representative olionucleotides in DU145 (human prostate cell line).

FIG. 24 depicts the structure of the olionucleotide IL8-1.

FIG. 25 depicts the structure of the olionucleotide IL8-3.

FIG. 26 demonstrates target inhibition of representative olionucleotides in BxPC3 (human pancreatic cancer cell line).

FIG. 27 demonstrates target inhibition of representative olionucleotides in A549 (human lung cancer cell line).

FIG. 28 depicts the structure of the olionucleotide KR1.

FIG. 29 depicts the structure of the olionucleotide KR2.

FIG. 30 depicts the structure of the olionucleotide KR0525.

FIG. 31 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 32 depicts the structure of the olionucleotide IL6.

FIG. 33 demonstrates target inhibition of representative olionucleotides in HCT-116 (human colorectal carcinoma).

FIG. 34 depicts the structure of the olionucleotide AKT4

FIG. 35 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 36 depicts the structure of the olionucleotide BC1.

FIG. 37 demonstrates target inhibition of representative olionucleotides in HCT-116 (human colorectal carcinoma).

FIG. 38 depicts the structure of the olionucleotide MEK1_(—)1.

FIG. 39 depicts the structure of the olionucleotide MEK1_(—)2.

FIG. 40 demonstrates target inhibition of representative olionucleotides in HCT-116 (human colorectal carcinoma).

FIG. 41 depicts the structure of the olionucleotide MEK2_(—)1.

FIG. 42 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 43 depicts the structure of the olionucleotide WNT1_(—)1.

FIG. 44 depicts the structure of the olionucleotide WNT1_(—)2.

FIG. 45 depicts the structure of the olionucleotide WNT1_(—)3.

FIG. 46 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 47 depicts the structure of the olionucleotide EZH2_(—)2.

FIG. 48 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 49 depicts the structure of the olionucleotide PD1.

FIG. 50 demonstrates target inhibition of representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 51 demonstrates target inhibition of representative olionucleotides in M14 (human melanoma cell line).

FIG. 52 demonstrates target inhibition of representative olionucleotides in NMuMG (a normal murine mouse mammary gland cell line).

FIG. 53 depicts the structure of the olionucleotide BL2.

FIG. 54 demonstrates target inhibition of representative olionucleotides in HCT-116 (human colorectal carcinoma).

FIG. 55 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 56 demonstrates target inhibition of representative olionucleotides in MDA-MB-231 a human breast cell line.

FIG. 57 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 58 depicts the structure of the olionucleotide CM7.

FIG. 59 depicts the structure of the olionucleotide CM12.

FIG. 60 depicts the structure of the olionucleotide CM13.

FIG. 61 depicts the structure of the olionucleotide CM14.

FIG. 62 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 63 depicts the structure of the olionucleotide TNF1.

FIG. 64 demonstrates target inhibition of representative olionucleotides in MCF7 (human mammary breast cell line).

FIG. 65 depicts the structure of the olionucleotide MIF1_(—)1.

FIG. 66 depicts the structure of the olionucleotide MIF1_(—)2.

FIG. 67 demonstrates that a representative oligonucleotide PC2 is capable of modulating target gene expression.

The figures are provided by way of example and are not intended to limit the scope of the present invention.

DETAILED DESCRIPTION OF THE INVENTION Definitions

To facilitate an understanding of the present invention, a number of terms and phrases are defined below:

As used herein, the term “wherein said chemotherapy agent is present at less than one half the standard dose” refers to a dosage that is less than one half (e.g., less than 50%, preferably less than 40%, even more preferably less than 10% and still more preferably less than 1%) of the minimum value of the standard dosage range used for dosing humans. In some embodiments, the standard dosage range is the dosage range recommended by the manufacturer. In other embodiments, the standard dosage range is the range utilized by a medical doctor in the field. In still other embodiments, the standard dosage range is the range considered the normal standard of care in the field. The particular dosage within the dosage range is determined, for example by the age, weight, and health of the subject as well as the type of cancer being treated.

As used herein, the term “under conditions such that expression of said gene is modulated” refers to conditions where an oligonucleotide of the present invention hybridizes to a gene) and modulates expression of the gene by at least 10%, preferably at least 25% relative to the level of transcription in the absence of the oligonucleotide. The present invention is not limited to the modulation of expression of a particular gene. Exemplary genes include, but are not limited to Survivin, Beclin-1, STAT3, HIF1A, IL-8, KRAS, MTTP, ApoC III, ApoB, IL-17, MMP2, FAP, P-selectin, IL-6, IL-23, AKT, CRAF, Beta Catenin, PCSK9, MEK1, MEK2, CD4, WNT1, Clusterin, NRAS, EZH2, HDAC1, and PD-1, TNFα, MIF1, TTR, HBV, HAMP, ERBB2, PARP1, ITGA4, APP, FGFR1, CD68, ALK, MSI2, JAK2, CCND1. As used herein, the term “under conditions such that transcription of said gene is modulated” refers to conditions where an oligonucleotide of the present invention hybridizes to a gene and modulates transcription of the gene by at least 10%, preferably at least 25% relative to the level of transcription in the absence of the oligonucleotide. The modulation of transcription of said gene may involve related genes. The present invention is not limited to the modulation of expression of a particular gene.

As used herein the term “expression” is the process whereby information from a gene is used in the synthesis of a functional gene product. These products may be proteins, but in non-protein coding genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or small nuclear RNA (snRNA) genes, the product is a functional RNA or transcript to generate the macromolecular machinery for gene expression. Gene expression may be modulated at several levels including transcription, RNA splicing, translation, and post-translational modification of a protein. The term may also be used against a viral gene and refer to mRNA synthesis from a RNA molecule (i.e. RNA replication). For instance, the genome of a negative-sense single-stranded RNA virus may serve as a template to translate the viral proteins for viral replication afterwards.

As used herein the term “transcription” is the first step of gene expression where a segment of DNA is copied into RNA by RNA polymerase to produce a transcript. If the gene transcribed encodes a protein, the result of transcription is messenger RNA (mRNA) and expressed to produce a protein. Alternatively, a transcribed gene may encode for non-coding RNA genes (e.g. such as microRNA etc.), ribosomal RNA, transfer RNA (tRNA), other components of the protein-assembly process, or other ribozymes.

As used herein the term “phenotype” describes the modulation of gene expression to define the properties of the expression give rise to the organism's phenotype. A phenotype is expressed by proteins that control the organism's characteristics or traits, such as its morphology, shape, development, biochemical or physiological properties, and products that act to catalyze cell signaling and metabolic pathways characterizing the organism.

As used herein the term “cell signaling” describes a complex system of signals or pathways that governs cellular activities and coordinates cell actions. A cell's ability to perceive and respond to its environment is processed through proteins involved in the cell signaling pathway.

As used herein the term “CG regions” are regions of DNA where cytosine and guanine nucleotides are enriched in the linear sequence of bases along the length of a gene. Generally CG or GC percentage that is greater than 50% with an observed-to-expected CpG ratio that is greater than 60%. CG regions of DNA are also where a cytosine nucleotide occurs next to a guanine nucleotide and may be referred to as “CpG” for “C phosphodiester bond G”. Generally cytosine bases in CpGs are methylated.

As used herein the term “CpG islands” are regions of the genome that have high GC content and higher concentration of CpG sites associated with the start of the gene, promoter regions or regions 5′ upstream of a gene start site. CpG islands are typically 300-3,000 base pairs in length. CpG islands are recognized to be hypomethylated. In most instances the CpG sites in the CpG islands are unmethylated and may be recognized by HpaII restriction site, CCGG.

As used herein the term “nuclease hypersensitive site” is a short region of chromatin and is detected by its super sensitivity to cleavage by DNase I and other various nucleases. The nucleosomal structure is less compact, increasing the availability of the DNA to binding by proteins, such as transcription factors and DNase I. Hypersensitive sites are found on chromatin of cells associated with genes and generally precede active promoters. When DNA is transcribed, 5′ hypersensitive sites appear before transcription begins, and the DNA sequences within the hypersensitive sites are required for gene expression. Hypersensitive sites may be generated as a result of the binding of transcription factors.

As used herein “cis-regulatory element” is a region of DNA or RNA that regulates the expression of genes located on that same molecule of DNA A cis-regulatory element may be located upstream of the coding sequence of the gene it controls (in the promoter region or even further upstream), in an intron, or downstream of the gene's coding sequence, in either the translated or the untranscribed region. A cis-regulatory element may be located in another gene other than the target gene in instances of chromosomal rearrangements.

As used herein “non-coding” refers to a linear sequence of DNA that does not contribute to an amino acid sequence of a protein.

As used herein “Trinucleotide repeat expansion” refers to a triplet repeat expansion of DNA bases that causes any type of disorder categorized as a trinucleotide repeat disorder. Generally, the larger the expansion the more likely they are to cause disease or increase the severity of disease. Trinucleotide repeat disorders represent genetic by trinucleotide repeat expansion, a kind of mutation where trinucleotide repeats in certain genes exceed the normal, stable threshold, which differs per gene.

As used herein, the term “under conditions such that growth of said cell is reduced” refers to conditions where an oligonucleotide of the present invention, when administered to a cell (e.g., a cancer) reduces the rate of growth of the cell by at least 10%, preferably at least 25%, even more preferably at least 50%, and still more preferably at least 90% relative to the rate of growth of the cell in the absence of the oligonucleotide.

As used herein, the term “under conditions such that the expression of said target is modulated” refers to conditions where an oligonucleotide of the present invention, when administered to a cell (e.g., a cancer or non cancer or immune cell) modulates the expression of the protein by at least 10%, preferably at least 25%, relative to basal expression in the absence of the oligonucleotide.

The term “epitope” as used herein refers to that portion of an antigen that makes contact with a particular antibody.

As used herein, the term “subject” refers to any animal (e.g., a mammal), including, but not limited to, humans, non-human primates, rodents, and the like, which is to be the recipient of a particular treatment. Typically, the terms “subject” and “patient” are used interchangeably herein in reference to a human subject.

As used herein, the terms “computer memory” and “computer memory device” refer to any storage media readable by a computer processor. Examples of computer memory include, but are not limited to, RAM, ROM, computer chips, digital video disc (DVDs), compact discs (CDs), hard disk drives (HDD), and magnetic tape.

As used herein, the term “computer readable medium” refers to any device or system for storing and providing information (e.g., data and instructions) to a computer processor. Examples of computer readable media include, but are not limited to, DVDs, CDs, hard disk drives, magnetic tape and servers for streaming media over networks.

As used herein, the term “Delta G” or “ΔG” is the change in Gibbs Free Energy (in units of kcal/mole) and is the net exchange of energy between the system and its environment and can be described by the equation ΔG=ΔH−T·ΔS. Where ΔH (Enthalpy) represents the total energy exchange between the system and its surrounding environment (in units of kcal/mole), ΔS (Entropy) represents the energy spent by the system to organize itself (in units of cal/K·mole). Generally speaking a spontaneous system favors a more random system not an ordered system. Finally, T represents the absolute temperature of the system and is in units Kelvin (Celsius +273.15). The change of free energy is equal to the sum of its enthalpy plus the product of the temperature and entropy of the system. A positive ΔG reaction is generally non-spontaneous while a negative value is spontaneous.

As used herein, the terms “processor” and “central processing unit” or “CPU” are used interchangeably and refer to a device that is able to read a program from a computer memory (e.g., ROM or other computer memory) and perform a set of steps according to the program.

As used herein, the term “non-human animals” refers to all non-human animals including, but are not limited to, vertebrates such as rodents, non-human primates, ovines, bovines, ruminants, lagomorphs, porcines, caprines, equines, canines, felines, ayes, etc. and and non-vertebrate animals such as drosophila and nematode. In some embodiments, “non-human animals” further refers to prokaryotes and viruses such as bacterial pathogens, fungal, viral pathogens. Non-human animals is used broadly here to also indicate plants and plant genomes, especially commercially valuable crops such as corn, soybean, cotton, the grasses and legumes including rice and alfalfa as well as commercial flowers, vegetables and trees including deciduous and evergreen.

As used herein, the term “nucleic acid molecule” refers to any nucleic acid containing molecule, including but not limited to, DNA or RNA. The term encompasses sequences that include any of the known base analogs of DNA and RNA including, but not limited to, 4-acetylcytosine, 8-hydroxy-N6-methyladenosine, aziridinylcytosine, pseudoisocytosine, 5-(carboxyhydroxylmethyl) uracil, 5-fluorouracil, 5-bromouracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, dihydrouracil, inosine, N6-isopentenyladenine, 1-methyladenine, 1-methylpseudouracil, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-methyladenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5′-methoxycarbonylmethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, oxybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, N-uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid, pseudouracil, queosine, 2-thiocytosine, and 2,6-diaminopurine.

The term “gene” refers to a nucleic acid (e.g., DNA) sequence that comprises coding sequences necessary for the production of a polypeptide, precursor, or RNA (e.g., rRNA, tRNA). The polypeptide can be encoded by a full length coding sequence or by any portion of the coding sequence so long as the desired activity or functional properties (e.g., enzymatic activity, ligand binding, signal transduction, immunogenicity, etc.) of the full-length or fragment are retained. The term also encompasses the coding region of a structural gene and the sequences located adjacent to the coding region on the 5′ ends for a distance of about 1 kb or more such that the gene corresponds to the length of the full-length mRNA. Sequences located 5′ of the coding region and present on the mRNA are referred to as 5′ non-translated sequences. Sequences located 3′ or downstream of the coding region and present on the mRNA are referred to as 3′ non-translated sequences. The term “gene” encompasses both cDNA and genomic forms of a gene. A genomic form or clone of a gene contains the coding region interrupted with non-coding sequences termed “introns” or “intervening regions” or “intervening sequences.” Introns are segments of a gene that are transcribed into nuclear RNA (hnRNA); introns may contain regulatory elements such as enhancers. Introns are removed or “spliced out” from the nuclear or primary transcript; introns therefore are absent in the messenger RNA (mRNA) transcript. The mRNA functions during translation to specify the sequence or order of amino acids in a nascent polypeptide.

As used herein, the term “heterologous gene” refers to a gene that is not in its natural environment. For example, a heterologous gene includes a gene from one species introduced into another species. A heterologous gene also includes a gene native to an organism that has been altered in some way (e.g., mutated, added in multiple copies, linked to non-native regulatory sequences, translocated, etc). Heterologous genes are distinguished from endogenous genes in that the heterologous gene sequences are typically joined to DNA sequences that are not found naturally associated with the gene sequences in the chromosome or are associated with portions of the chromosome not found in nature (e.g., genes expressed in loci where the gene is not normally expressed).

As used herein, the term “gene expression” refers to the process of converting genetic information encoded in a gene into RNA (e.g., mRNA, rRNA, tRNA, or snRNA) through “transcription” of the gene (i.e., via the enzymatic action of an RNA polymerase), and for protein encoding genes, into protein through “translation” of mRNA. Gene expression can be regulated at many stages in the process. “Up-regulation” or “activation” refers to regulation that increases the production of gene expression products (i.e., RNA or protein), while “down-regulation” or “repression” refers to regulation that decrease production. “Modulation” refers to regulation that is altered. Molecules (e.g., transcription factors) that are involved in up-regulation or down-regulation are often called “activators” and “repressors or suppressors,” respectively.

In addition to containing introns, genomic forms of a gene may also include sequences located on both the 5′ and 3′ end of the sequences that are present on the RNA transcript. These sequences are referred to as “flanking” sequences or regions (these flanking sequences are located 5′ or 3′ to the non-translated sequences present on the mRNA transcript). The 5′ flanking region may contain regulatory sequences such as promoters and enhancers that control or influence the transcription of the gene. The 3′ flanking region may contain sequences that direct the termination of transcription, post-transcriptional cleavage and polyadenylation.

The term “wild-type” refers to a gene or gene product isolated from a naturally occurring source. A wild-type gene is that which is most frequently observed in a population and is thus arbitrarily designed the “normal” or “wild-type” form of the gene. In contrast, the term “modified” or “mutant” refers to a gene or gene product that displays modifications in sequence and/or functional properties (i.e., altered characteristics) or phenotype when compared to the wild-type gene or gene product. It is noted that naturally occurring mutants can be isolated; these are identified by the fact that they have altered characteristics (including altered nucleic acid sequences) when compared to the wild-type gene or gene product.

As used herein, the terms “nucleic acid molecule encoding,” “DNA sequence encoding,” and “DNA encoding” refer to the order or sequence of deoxyribonucleotides along a strand of deoxyribonucleic acid. The order of these deoxyribonucleotides determines the order of amino acids along the polypeptide (protein) chain. The DNA sequence thus codes for the amino acid sequence.

As used herein, the terms “an oligonucleotide having a nucleotide sequence encoding a gene” and “polynucleotide having a nucleotide sequence encoding a gene,” means a nucleic acid sequence comprising the coding region of a gene or in other words the nucleic acid sequence that encodes a gene product. The coding region may be present in a cDNA, genomic DNA or RNA form. When present in a DNA form, the oligonucleotide or polynucleotide may be single-stranded (i.e., the sense strand) or double-stranded. Suitable control elements such as enhancers/promoters, splice junctions, polyadenylation signals, etc. may be placed in close proximity to the coding region of the gene if needed to permit proper initiation of transcription and/or correct processing of the primary RNA transcript. Alternatively, the coding region utilized in the expression vectors of the present invention may contain endogenous enhancers/promoters, splice junctions, intervening sequences, polyadenylation signals, etc. or a combination of both endogenous and exogenous control elements.

As used herein, the term “oligonucleotide,” refers to a short length of single-stranded polynucleotide chain. Oligonucleotides are typically less than 200 residues long (e.g., between 8 and 100), however, as used herein, the term is also intended to encompass longer polynucleotide chains (e.g., as large as 5000 residues). Oligonucleotides are often referred to by their length. For example a 24 residue or base oligonucleotide is referred to as a “24-mer”. Oligonucleotides can form secondary and tertiary structures by self-hybridizing or by hybridizing to other polynucleotides. Such structures can include, but are not limited to, duplexes, hairpins, cruciforms, bends, and triplexes.

In some embodiments, oligonucleotides are “DNAi or DNA interference (DNAi).” As used herein, the term “DNAi” or refers to an oligonucleotide that hybridizes to region 5′ upstream of the transcription start site of a gene. In some embodiments, the hybridization of the DNAi or DNAi to the promoter modulates expression of the gene.

As used herein, the terms “complementary” or “complementarity” are used in reference to polynucleotides (i.e., a sequence of nucleotides) related by the base-pairing rules. For example, for the sequence “A-G-T,” is complementary to the sequence “T-C-A.” Complementarity may be “partial,” in which only some of the nucleic acids' bases are matched according to the base pairing rules. Or, there may be “complete” or “total” or “100 percent” complementarity between the nucleic acids. The degree of complementarity between nucleic acid strands has significant effects on the efficiency and strength of hybridization between nucleic acid strands. The degree of complementarity is also defined the “native” sequence rather than having a mismatch. This is of particular importance in amplification reactions, as well as detection methods that depend upon binding between nucleic acids.

As used herein, the term “completely complementary,” for example when used in reference to an oligonucleotide of the present invention refers to an oligonucleotide where all of the nucleotides are complementary to a target sequence (e.g., a gene).

As used herein, the term “partially complementary,” for example when used in reference to an oligonucleotide of the present invention, refers to an oligonucleotide where at least one nucleotide is not complementary to the target sequence. Preferred partially complementary oligonucleotides are those that can still hybridize to the target sequence under physiological conditions. The term “partially complementary” refers to oligonucleotides that have regions of one or more non-complementary nucleotides both internal to the oligonucleotide or at either end. Oligonucleotides with mismatches at the ends may still hybridize to the target sequence.

The term “homology” refers to a degree of complementarity. There may be partial homology or complete homology (i.e., identity). A partially complementary sequence is a nucleic acid molecule that at least partially inhibits a completely complementary nucleic acid molecule from hybridizing to a target nucleic acid is “substantially homologous.” The inhibition of hybridization of the completely complementary sequence to the target sequence may be examined using a hybridization assay (Southern or Northern blot, solution hybridization and the like) under conditions of low stringency. A substantially homologous sequence or probe will compete for and inhibit the binding (i.e., the hybridization) of a completely homologous nucleic acid molecule to a target under conditions of low stringency. This is not to say that conditions of low stringency are such that non-specific binding is permitted; low stringency conditions require that the binding of two sequences to one another be a specific (i.e., selective) interaction. The absence of non-specific binding may be tested by the use of a second target that is substantially non-complementary (e.g., less than about 30% identity); in the absence of non-specific binding the probe will not hybridize to the second non-complementary target.

When used in reference to a double-stranded nucleic acid sequence such as a cDNA or genomic clone, the term “substantially homologous” refers to any probe that can hybridize to either or both strands of the double-stranded nucleic acid sequence under conditions of low stringency as described above.

A gene may produce multiple RNA species that are generated by differential splicing of the primary RNA transcript. cDNAs that are splice variants of the same gene will contain regions of sequence identity or complete homology (representing the presence of the same exon or portion of the same exon on both cDNAs) and regions of complete non-identity (for example, representing the presence of exon “A” on cDNA 1 wherein cDNA 2 contains exon “B” instead). Because the two cDNAs contain regions of sequence identity they will both hybridize to a probe derived from the entire gene or portions of the gene containing sequences found on both cDNAs; the two splice variants are therefore substantially homologous to such a probe and to each other.

When used in reference to a single-stranded nucleic acid sequence, the term “substantially homologous” refers to any probe that can hybridize (i.e., it is the complement of) the single-stranded nucleic acid sequence under conditions of low stringency as described above.

As used herein, the term “hybridization” is used in reference to the pairing of complementary nucleic acids. Hybridization and the strength of hybridization (i.e., the strength of the association between the nucleic acids) is impacted by such factors as the degree of complementary between the nucleic acids, stringency of the conditions involved, the Tm of the formed hybrid, and the G:C or C:G ratio within the nucleic acids. An oligonucleotide is a single molecule that contains a covalent bond linking each nucleotide and often pairing of complementary nucleic acids within its structure is said to be “self-hybridized” or having secondary structure.

As used herein the term “secondary structure” means a single molecule that contains a pairing of complementary nucleic acids within its structure that contributes to a two dimensional bend in said molecule.

As used herein, the term “linear section” refers to molecules with secondary structures wherein those secondary structures have regions of DNA that are not paired in a secondary manner they only have one covalent bond to the next oligonucleotide rather than both a bond and a pairing of complementary nucleic acids as one finds in regions having secondary structure.”

As used herein, the term “nuclease hypersensitive region” refers to regions of the target gene that are susceptible to oligonucleotide binding.

As used herein, the term “Tm” is used in reference to the “melting temperature.” The melting temperature is the temperature at which a population of double-stranded nucleic acid molecules becomes half dissociated into single strands. The equation for calculating the Tm of nucleic acids is well known in the art. As indicated by standard references, a simple estimate of the Tm value may be calculated by the equation: Tm=81.5+0.41 (% G+C), when a nucleic acid is in aqueous solution at 1 M NaCl (See e.g., Anderson and Young, Quantitative Filter Hybridization, in Nucleic Acid Hybridization [1985]). Other references include more sophisticated computations that take structural as well as sequence characteristics into account for the calculation of Tm. The process of hybridization and dissociation is complex and highly dynamic and at the Tm, double strands are constantly formed and broken up, resulting in multiple interactions over time. The formation of secondary structures within an oligonucleotide may influence Tm.

As used herein the term “stringency” is used in reference to the conditions of temperature, ionic strength, and the presence of other compounds such as organic solvents, under which nucleic acid hybridizations are conducted. Under “low stringency conditions” a nucleic acid sequence of interest will hybridize to its exact complement, sequences with single base mismatches, closely related sequences (e.g., sequences with 90% or greater homology), and sequences having only partial homology (e.g., sequences with 50-90% homology). Under “medium stringency conditions,” a nucleic acid sequence of interest will hybridize only to its exact complement, sequences with single base mismatches, and closely relation sequences (e.g., 90% or greater homology). Under “high stringency conditions,” a nucleic acid sequence of interest will hybridize only to its exact complement, and (depending on conditions such a temperature) sequences with single base mismatches. In other words, under conditions of high stringency the temperature can be raised so as to exclude hybridization to sequences with single base mismatches.

“High stringency conditions” when used in reference to nucleic acid hybridization comprise conditions equivalent to binding or hybridization at 42° C. in a solution consisting of 5×SSPE (43.8 g/l NaCl, 6.9 g/l NaH2PO4 H2O and 1.85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.5% SDS, 5×Denhardt's reagent and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 0.1×SSPE, 1.0% SDS at 42° C. when a probe of about 500 nucleotides in length is employed.

“Medium stringency conditions” when used in reference to nucleic acid hybridization comprise conditions equivalent to binding or hybridization at 42° C. in a solution consisting of 5×SSPE (43.8 g/l NaCl, 6.9 g/l NaH2PO4 H2O and 1.85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.5% SDS, 5×Denhardt's reagent and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 1.0×SSPE, 1.0% SDS at 42° C. when a probe of about 500 nucleotides in length is employed.

“Low stringency conditions” comprise conditions equivalent to binding or hybridization at 42° C. in a solution consisting of 5×SSPE (43.8 g/l NaCl, 6.9 g/l NaH2PO4 H2O and 1.85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.1% SDS, 5×Denhardt's reagent [50×Denhardt's contains per 500 ml: 5 g Ficoll (Type 400, Pharamcia), 5 g BSA (Fraction V; Sigma)] and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 5×SSPE, 0.1% SDS at 42° C. when a probe of about 500 nucleotides in length is employed.

The present invention is not limited to the hybridization of probes of about 500 nucleotides in length. The present invention contemplates the use of probes between approximately 8 nucleotides up to several thousand (e.g., at least 5000) nucleotides in length. One skilled in the relevant understands that stringency conditions may be altered for probes of other sizes (See e.g., Anderson and Young, Quantitative Filter Hybridization, in Nucleic Acid Hybridization [1985] and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, NY [1989]).

One skilled in the art would know numerous equivalent conditions may be employed to create low stringency conditions; factors such as the length and nature (DNA, RNA, base composition) of the probe and nature of the target (DNA, RNA, base composition, present in solution or immobilized, etc.) and the concentration of the salts and other components (e.g., the presence or absence of formamide, dextran sulfate, polyethylene glycol) are considered and the hybridization solution may be varied to generate conditions of low stringency hybridization different from, but equivalent to, the above listed conditions. In addition, the art knows conditions that promote hybridization under conditions of high stringency (e.g., increasing the temperature of the hybridization and/or wash steps, the use of formamide in the hybridization solution, etc.) (see definition above for “stringency”).

As used herein, the term “physiological conditions” refers to specific stringency conditions that approximate or are conditions inside an animal (e.g., a human). Exemplary physiological conditions for use in vitro include, but are not limited to, 37° C., 95% air, 5% CO2, commercial medium for culture of mammalian cells (e.g., DMEM media available from Gibco, Md.), 5-10% serum (e.g., calf serum or horse serum), additional buffers, and optionally hormone (e.g., insulin and epidermal growth factor).

The term “isolated” when used in relation to a nucleic acid, as in “an isolated oligonucleotide” or “isolated polynucleotide” refers to a nucleic acid sequence that is identified and separated from at least one component or contaminant with which it is ordinarily associated in its natural source. Isolated nucleic acid is such present in a form or setting that is different from that in which it is found in nature. In contrast, non-isolated nucleic acids as nucleic acids such as DNA and RNA found in the state they exist in nature. For example, a given DNA sequence (e.g., a gene) is found on the host cell chromosome in proximity to neighboring genes; RNA sequences, such as a specific mRNA sequence encoding a specific protein, are found in the cell as a mixture with numerous other mRNAs that encode a multitude of proteins. However, isolated nucleic acid encoding a given protein includes, by way of example, such nucleic acid in cells ordinarily expressing the given protein where the nucleic acid is in a chromosomal location different from that of natural cells, or is otherwise flanked by a different nucleic acid sequence than that found in nature. The isolated nucleic acid, oligonucleotide, or polynucleotide may be present in single-stranded or double-stranded form. When an isolated nucleic acid, oligonucleotide or polynucleotide is to be utilized to express a protein, the oligonucleotide or polynucleotide will contain at a minimum the sense or coding strand (i.e., the oligonucleotide or polynucleotide may be single-stranded), but may contain both the sense and anti-sense strands (i.e., the oligonucleotide or polynucleotide may be double-stranded).

As used herein, the term “purified” or “to purify” refers to the removal of components (e.g., contaminants) from a sample. For example, antibodies are purified by removal of contaminating non-immunoglobulin proteins; they are also purified by the removal of immunoglobulin that does not bind to the target molecule. The removal of non-immunoglobulin proteins and/or the removal of immunoglobulins that do not bind to the target molecule results in an increase in the percent of target-reactive immunoglobulins in the sample. In another example, recombinant polypeptides are expressed in bacterial host cells and the polypeptides are purified by the removal of host cell proteins; the percent of recombinant polypeptides is thereby increased in the sample.

“Amino acid sequence” and terms such as “polypeptide” or “protein” are not meant to limit the amino acid sequence to the complete, native amino acid sequence associated with the recited protein molecule.

The term “native protein” as used herein to indicate that a protein does not contain amino acid residues encoded by vector sequences; that is, the native protein contains only those amino acids found in the protein as it occurs in nature. A native protein may be produced by recombinant means or may be isolated from a naturally occurring source.

The term “mutant protein” as used herein to indicate that a protein containing a change in amino acid residues encoded by vector sequences that renders altered function or implicated in disease; that is, the mutant protein contains only those amino acids found in the protein as it occurs in nature. A mutant protein may be produced by recombinant means or may be isolated from a naturally occurring source

As used herein the term “portion” when in reference to a protein (as in “a portion of a given protein”) refers to fragments of that protein. The fragments may range in size from four amino acid residues to the entire amino acid sequence minus one amino acid.

The term “Southern blot,” refers to the analysis of DNA on agarose or acrylamide gels to fractionate the DNA according to size followed by transfer of the DNA from the gel to a solid support, such as nitrocellulose or a nylon membrane. The immobilized DNA is then probed with a labeled probe to detect DNA species complementary to the probe used. The DNA may be cleaved with restriction enzymes prior to electrophoresis. Following electrophoresis, the DNA may be partially depurinated and denatured prior to or during transfer to the solid support. Southern blots are a standard tool of molecular biologists (J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, NY, pp 9.31-9.58 [1989]).

The term “Northern blot,” as used herein refers to the analysis of RNA by electrophoresis of RNA on agarose gels to fractionate the RNA according to size followed by transfer of the RNA from the gel to a solid support, such as nitrocellulose or a nylon membrane. The immobilized RNA is then probed with a labeled probe to detect RNA species complementary to the probe used. Northern blots are a standard tool of molecular biologists (J. Sambrook, et al., supra, pp 7.39-7.52 [1989]).

The term “Western blot” refers to the analysis of protein(s) (or polypeptides) immobilized onto a support such as nitrocellulose or a membrane. The proteins are run on acrylamide gels to separate the proteins, followed by transfer of the protein from the gel to a solid support, such as nitrocellulose or a nylon membrane. The immobilized proteins are then exposed to antibodies with reactivity against an antigen of interest. The binding of the antibodies may be detected by various methods, including the use of radiolabeled antibodies.

As used herein, the term “cell culture” refers to any in vitro culture of cells. Included within this term are continuous cell lines (e.g., with an immortal phenotype), primary cell cultures, transformed cell lines, finite cell lines (e.g., non-transformed cells), and any other cell population maintained in vitro.

As used, the term “eukaryote” refers to organisms distinguishable from “prokaryotes.” It is intended that the term encompass all organisms with cells that exhibit the usual characteristics of eukaryotes, such as the presence of a true nucleus bounded by a nuclear membrane, within which lie the chromosomes, the presence of membrane-bound organelles, and other characteristics commonly observed in eukaryotic organisms. Thus, the term includes, but is not limited to such organisms as fungi, protozoa, and animals (e.g., humans).

As used herein, the term “in vitro” refers to an artificial environment and to processes or reactions that occur within an artificial environment. In vitro environments can consist of, but are not limited to, test tubes and cell culture. The term “in vivo” refers to the natural environment (e.g., an animal or a cell) and to processes or reaction that occur within a natural environment.

The terms “test compound” and “candidate compound” refer to any chemical entity, pharmaceutical, drug, and the like that is a candidate for use to treat or prevent a disease, illness, sickness, disorder of bodily function (e.g., cancer or non-cancer disease) or disrupt a system (e.g. cell culture). Test compounds comprise both known and potential therapeutic compounds. A test compound can be determined to be therapeutic by screening using the screening methods of the present invention. In some embodiments of the present invention, test compounds include antisense compounds.

As used herein, the term “known chemotherapeutic agents” refers to compounds known to be useful in the treatment of disease (e.g., cancer). Exemplary chemotherapeutic agents affective against cancer include, but are not limited to, daunorubicin, dactinomycin, doxorubicin, bleomycin, mitomycin, nitrogen mustard, chlorambucil, melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine (CA), 5-fluorouracil (5-FU), floxuridine (5-FUdR), methotrexate (MTX), colchicine, vincristine, vinblastine, etoposide, teniposide, cisplatin, lenolamide, and diethylstilbestrol (DES).

As used herein, the term “sample” is used in its broadest sense. In one sense, it is meant to include a specimen or culture obtained from any source, as well as biological and environmental samples. Biological samples may be obtained from animals (including humans) and encompass fluids, solids, tissues, and gases. Biological samples include blood products, such as plasma, serum and the like. Environmental samples include environmental material such as surface matter, soil, water, crystals and industrial samples. Such examples are not however to be construed as limiting the sample types applicable to the present invention.

“Hot Zones” in some embodiments, are regions within the promoter region of an oncogene are further defined as preferred regions for hybridization of oligonucleotides. In some embodiments, these preferred regions are referred to as “hot zones.” In some preferred embodiments, hot zones are defined based on oligonucleotide compounds that are demonstrated to be effective (see above section on oligonucleotides) and those that are contemplated to be effective based on the preferred criteria for oligonucleotides described above. Preferred hot zones encompass 20 bp upstream and downstream of each compound included in each hot zone and have at least 1 CG or more within an increment of 40 bp further upstream or downstream of each compound. In preferred embodiments, hot zones encompass a maximum of 100 bp upstream and downstream of each oligonucleotide compound included in the hot zone. In additional embodiments, hot zones are defined at beginning regions of each promoter. These hot zones are defined either based on effective sequence(s) or contemplated sequences and have a preferred maximum length of 1000 bp. Based on the above described criteria, exemplary hot zones were designed. Specific hot zones are described in the examples.

Combination and Single-Agent Therapy Using this DNAi Technology.

We present and define the following disease conditions as exemplary of, but not limited to, those that are potentially treatable with the DNAi therapeutic(s) described herein. Treatment of these disease entities may occur with single-agent DNAi therapy or DNAi therapy in combination with one or more therapeutics used to treat the conditions.

Cardiovascular Disease

Treating cardiovascular disease involves opening narrowed arteries, correcting abnormalities associated with irregular heartbeats and dysfunctional heart muscle or valves, reducing high blood pressure and high lipid levels, and amending imbalances in clotting that causes symptoms of pain and discomfort. Inventions may include: medical devices, dyslipidemics, antithrombotics, anticoagulants, anti-platelets, antihypertensives, anti-inflammatory, antihypertrophics, diuretics, anti-anginal, channel blockers, anti-restenosis agents, anti-atherosclerotics, anti-arrhythmics, enzyme inhibitors, and complement inhibitors.

Antianginals

The heart muscle works continuously and requires a constant supply of nutrients and oxygen. Those nutrients and oxygen are carried to the heart muscle in the blood. The chest pain known as angina can occur when there is an insufficient supply of blood, and consequently of oxygen, to the heart muscle. There are several types of antianginal medications. These include beta blockers (acebutolol, atenolol, betaxolol, bisoprolol, labetalol, metoprolol, nadolol, pindolol, propranolol, timolol), calcium channel blockers (diltiazem, nifedipine, verapamil), and vasodilators (nitroglycerin, isosorbide dinitrate). These drugs act by increasing the amount of oxygen that reaches the heart muscle.

Antiarrhythmics

Antiarrhythmics are used when the heart does not beat rhythmically or smoothly (a condition called arrhythmia), its rate of contraction must be regulated. Antiarrhythmic drugs (disopyramide, mexiletine, procainamide, propranolol, amiodarone, tocainide) prevent or alleviate arrhythmias by altering nerve impulses in the heart. Anticoagulants are used when clots develop on the interior wall of an artery block blood flow.

Antihyperlipidemics

Medications for treating atherosclerosis, or hardening of the arteries, act to reduce the serum levels of cholesterol and triglycerides, which form plaques on the walls of arteries. The following drug classes are used to treat high cholesterol or high lipid levels: HMG CoA reductase inhibitors (atorvastatin, simvastatin, lovastatin, and rosuvastatin, fluvastatin, pravastatin), fibrates (fenofibrate, gemfibrozil), bile acid sequestrants (cholestyramine, colestipol, and colesevelam), niacins (niacin, Vit B3, nicotinic acid), and cholesterol absorption inhibitors (ezetimide), or drug combinations of these classes.

Antihypertensives

High blood pressure is caused when the pressure of the blood against the walls of the blood vessels is higher than what is considered normal. High blood pressure, or hypertension, eventually causes damage to the brain, eyes, heart, or kidneys. Several different drug actions produce an antihypertensive effect. Some drugs block nerve impulses that cause arteries to constrict; others slow the heart rate and decrease its force of contraction; still others reduce the amount of a certain hormone in the blood that causes blood pressure to rise. The effect of any of these medications is to reduce blood pressure. The mainstay of antihypertensive therapy is often a diuretic, a drug that reduces body fluids. Examples of antihypertensive drugs include beta blockers, calcium channel blockers, ACE (angiotensin-converting enzyme) inhibitors (including benazepril, captopril, enalapril, lisinopril, and quinapril), and the agents valsartan, losartan, prazosin, and terazosin.

Antiplatelets

Antilatelet drugs alter the platelet activation at the site of vascular damage crucial to the development of arterial thrombosis. Aspirin irreversibly inhibits the enzyme COX, resulting in reduced platelet production of TXA2 (thromboxane—powerful vasoconstrictor that lowers cyclic AMP and initiates the platelet release reaction). Dipyridamole inhibits platelet phosphodiesterase, causing an increase in cyclic AMP with potentiation of the action of PGI2-—opposes actions of TXA2. Clopidogrel (Plavix) affects the ADP-dependent activation of IIb/IIIa complex. Glycoprotein IIb/IIIa receptor antagonists block a receptor on the platelet for fibrinogen and von Willebrand factor and include for example, abciximab eptifibatide and tirofiban. Epoprostenol is a prostacyclin that is used to inhibit platelet aggregation during renal dialysis (with or without heparin) and is also used in primary pulmonary hypertension.

Antithrombotics

An antithrombotic agent is a drug that reduces thrombus formation. These include plasminogen activators: Alteplase, Reteplase, Tenecteplase, Saruplase, Urokinase, Anistreplase, Monteplase, Streptokinase, other serine endopeptidases (Ancrod, Brinase, Fibrinolysin)

Beta Blockers

Beta-blocking medications block the response of the heart and blood vessels to nerve stimulation, thereby slowing the heart rate and lowering blood pressure. They are used in the treatment of a wide range of diseases, including angina, high blood pressure, migraine headaches, arrhythmias, and glaucoma. Metoprolol and propranolol are common beta blockers.

Calcium Channel Blockers

Calcium channel blockers (diltiazem, nifedipine, verapamil) are used for the prevention of angina (chest pain). Verapamil is also useful in correcting certain arrhythmias (heartbeat irregularities) and lowering blood pressure. This group of drugs is thought to prevent angina and arrhythmias and lower blood pressure by blocking or slowing calcium flow into muscle cells, which results in vasodilation (widening of the blood vessels) and greater oxygen delivery to the heart muscle.

Cardiac Glycosides

Cardiac glycosides include drugs that are derived from digitalis (digoxin is an example). This type of drug slows the rate of the heart but increases its force of contraction. Cardiac glycosides act as both heart depressants and stimulants: They may be used to regulate irregular heart rhythm or to increase the volume of blood pumped by the heart in heart failure.

Diuretics

Diuretic drugs, such as chlorothiazide, chlorthalidone, furosemide, hydrochlorothiazide, and spironolactone, promote the loss of water and salt from the body to lower blood pressure or increase the diameter of blood vessels. Antihypertensive medications cause the body to retain salt and water and are often used concurrently with diuretics. Most diuretics act directly on the kidneys, but there are different types of diuretics, each with different actions. This allows therapy for high blood pressure to be adjusted to meet the needs of individual patients.

Thiazide diuretics, such as chlorothiazide, chlorthalidone, and hydrochlorothiazide, are the most commonly prescribed and generally well tolerated as once or twice a day pills. A major drawback of thiazide diuretics is that they often deplete the body of potassium and therefore compensated with potassium supplements. Loop diuretics, such as furosemide, act more vigorously than thiazide diuretics. (Loop refers to the structures in the kidneys on which these specific diuretic medications act.) Loop diuretics promote more water loss than thiazide diuretics but they also deplete more potassium from the body. Potassium sparing diuretics are also used treat heart failure and high blood pressure and include amiloride, spironolactone, and triamterene. Generally drug combinations of amiloride and hydrochlorothiazide, spironolactone and hydrochlorothiazide, and triamterene and hydrochlorothiazide are used to enhance the antihypertensive effect and reduce potassium loss.

Vasodilators

Vasodilating medications cause the blood vessels to dilate, or widen. Some of the antihypertensive medications, such as hydralazine and prazosin, lower blood pressure by dilating the arteries or veins. Other vasodilating medicines are used in the treatment of stroke and diseases that are characterized by poor blood circulation. Ergoloid mesylates, for example, are used to reduce the symptoms of senility by increasing the flow of oxygen-rich blood to the brain.

Metabolic Disease (Diabetes)

Diabetes is usually a lifelong or chronic disease caused by high levels of sugar in the blood. Insulin is a produced by the pancreas to control blood sugar and diabetes can be caused by too little insulin, resistance to insulin, or both. There are several types of diabetes. (1) Type 1 diabetes can occur at any age, but it is most often diagnosed in children, teens, or young adults. It is caused by the destruction of islet cells in the pancreas resulting in little or no insulin thereby requiring daily injections of insulin. (2) Type 2 diabetes results from insulin resistance and relative insulin deficiency. Obesity is thought to be the primary cause of Type 2 diabetes in those genetically predisposed. (3) Gestational diabetes is high blood sugar that develops at any time during pregnancy in a woman who does not have diabetes.

The following treatments for diabetes include: insulin, biguanides (metformin), suphonylureas, nonsulfonylurea secretagogues, meglitinides/prandial glucose regulatory/glinides, alpha-glucosidase inhibitors, thiazolidineione/glitazones, glucagon-like peptide-1 analog, amylin analogues, and dipeptidyl peptidase-4 inhibitors.

Metformin is generally recommended as a first line treatment. When metformin is not sufficient another class is added.

Sulfonylureas lower blood sugar by stimulating the pancreas to release more insulin. The first drugs of this type that were developed—Dymelor (acetohexamide), Diabinese (chlorpropamide), Orinase (tolbutamide), and Tolinase (tolazamide)—are not as widely used since they tend to be less potent and shorter-acting drugs than the newer sulfonylureas. They include Glucotrol (glipizide), Glucotrol XL (extended release), DiaBeta (glyburide), Micronase (glyburide), Glynase PresTab (glyburide), and Amaryl (glimepiride). These drugs can cause a decrease in the hemoglobin A1c (HbA1c) of up to 1%-2%. Biguanides improve insulin's ability to move sugar into cells especially into the muscle cells and prevent the liver from releasing stored sugar. Biguanides are counterindicated in people who have kidney damage or heart failure because of the risk of precipitating a severe build-up of lactic acid (called lactic acidosis) in these patients. Biguanides can decrease the HbA1c 1%-2%. An example includes metformin (Glucophage, Glucophage XR, Riomet, Fortamet, and Glumetza).

Thiazolidinediones improve insulin's effectiveness (improving insulin resistance) in muscle and in fat tissue. They lower the amount of sugar released by the liver and make fat cells more sensitive to the effects of insulin. Actos (pioglitazone) and Avandia (rosiglitazone) are the two drugs of this class. A decrease in the HbA1c of 1%-2% can be seen with this class of oral diabetes medications. Thiazolidinediones should used with caution in people with heart failure. Avandia is restricted for use in new patients only if they are uncontrolled on other medications and are unable to take Actos.

Alpha-glucosidase inhibitors include Precose (acarbose) and Glyset (miglitol). These drugs block enzymes that help digest starches, slowing the rise in blood sugar. These diabetes pills may cause diarrhea or gas. They can lower hemoglobin A1c by 0.5%-1%.

Meglitinides include Prandin (repaglinide) and Starlix (nateglinide). These diabetes medicines lower blood sugar by stimulating the pancreas to release more insulin. The effects of these drugs are glucose-dependent, with high blood sugar inducing insulin release, which is unlike the action of sulfonylureas which cause insulin release, regardless of glucose levels, and can lead to hypoglycemia.

Dipeptidyl peptidase IV (DPP-IV) inhibitors include Januvia (sitagliptin), Nesina (alogliptin), Onglyza (saxagliptin), Galvus (vildagliptin) and Tradjenta (linagliptin). The DPP-IV inhibitors work to lower blood sugar in patients with type 2 diabetes by increasing insulin secretion from the pancreas and reducing sugar production. These diabetes pills increase insulin secretion when blood sugars are high. They also signal the liver to stop producing excess amounts of sugar. DPP-IV inhibitors control sugar without causing weight gain. The medication may be taken alone or with other medications such as metformin.

Glucagon-Like Peptide Analogs and Agonists

Glucagon-like peptide (GLP) agonists bind to a membrane GLP receptor. As a consequence, insulin release from the pancreatic beta cells is increased. Examples of this class include Exenatide (also Exendin-4, marketed as Byetta). Exenatide is not an analogue of GLP but rather a GLP agonist. Typical reductions in A1C values are 0.5-1.0%. Liraglutide, a once-daily human analogue (97% homology), has been developed by Novo Nordisk under the brand name Victoza. Taspoglutide is presently in Phase III clinical trials with Hoffman-La Roche.

Alpha-glucosidase inhibitors (Acarbose, Miglitol, Voglibose), amylin analogues (Pramlintide), SGLT2 inhibitors (Canagliflozin, Dapagliflozin, Empaliflozin, Remogliflozin, Sergliflozin) and others (Benfluorex, Tolrestat)

Combination agents are the combination of two medications in one tablet and include the following examples: Glucovance, which combines glyburide (a sulfonylurea) and metformin, Metaglip, which combines glipizide (a sulfonylurea) and metformin, and Avandamet which utilizes both metformin and rosiglitazone (Avandia). Kazano (alogliptin and metformin) and Oseni (alogliptin plus pioglitazone) are other examples.

Eye Disorders

Ocular Bacterial Infection. Antibiotics are generally used to treat, or sometimes to prevent a bacterial eye infection. Examples of common antibiotics used in the eye are sulfacetamide, erythromycin, gentamicin, tobramycin, ciprofloxacin and ofloxacin.

Ocular Inflammatory reaction. Anti-inflammatories reduce inflammation, which in the eye is usually manifest by pain, redness, light sensitivity and sometimes blurred vision. Anti-inflammatories can be either glucocorticoids/corticosteroids or NSAIDs. Corticosteroids are very effective anti-inflammatories for a wide variety of eye problems including all disorders associated with systemic inflammatory reactions (Reiter's syndrome, xerostomia, etc.). Common corticosteroids include: Prednisolone, Fluorometholone and Dexamethasone. Non-steroidal anti-inflammatories reduce the production of pro-inflammatory factors such as prostaglandins. Common NSAIDs include: Diclofenac, Ketorolac and Flurbiprofen.

Glaucoma. Glaucoma is a disorder of regulation of intraocular pressure. Glaucoma medications all attempt to reduce this pressure to prevent damage to the optic nerve resulting in loss of vision. These medications may lower pressure by decreasing the amount of fluid produced in the eye, by increasing the amount of fluid exiting through the eye's natural drain, or by providing additional pathways for fluid to leave the eye. More than one glaucoma medication is used simultaneously, as these effects can combine to lower pressure further than possible with a single medication. These medications are listed by class:

BETA-BLOCKERS: Timolol, Metipranolol, Carteolol, Betaxolol, Levobunolol ALPHA AGONISTS: Brimonidine, Iopidine PROSTAGLANDIN ANALOGUES: Latanoprost CARBONIC ANHYDRASE INHIBITORS: Dorzolamide CHOLINERGIC AGONISTS: Pilocarpine, Carbachol ADENERGIC AGONISTS, Epinephrine, Dipivefrin

Ocular Viral Infection

Used primarily in treating herpes virus infections of the eye, antiviral eye medications may be used in conjunction with oral medications for elimination the virus. The most common type of antiviral is triflurthymidine. Other topical anti-virals include adenine arabinoside and idoxuridine.

Allergic Reaction

All anti-allergy topicals decrease the effects of histamine, a factor that mediates, the inflammatory reaction. Common anti-allergy medicines include livostin, patanol, Cromolyn and alomide.

Infectious Diseases

Aminoglycosides. This class of antibiotics is used to treat infections caused by Gram-negative bacteria, such as Escherichia coli and Klebsiella, particularly Pseudomonas aeruginosa. This class is also effective against Aerobic bacteria (but not obligate/facultative anaerobes) and in the treatment of tularemia. The mechanism of action includes binding to the bacterial 30S ribosome/ribosomal subunit (some work by binding to the 50S subunit), inhibiting the translocation of the peptidyl-tRNA from the A-site to the P-site and also causing misreading of mRNA, leaving the bacterium unable to synthesize proteins vital to its growth. Possible toxicities include hearing loss, vertigo and nephrotoxicity. Examples of aminoglycosides include Amikacin, Gentamicin, Kanamycin, Neomycin, Netilmicin, Tobramycin, Paromomycin, Spectinomycin.

Ansamycins. Used as anti-tumor antibiotics and for treatment of traveler's diarrhea caused by E. coli. Examples include Geldanamycin, Herbimycin, and Rifaximin.

Carbacephem. This class prevents bacterial cell division by inhibiting cell wall synthesis. An example is Loracarbef.

Carbapenem. This class works by inhibiting cell wall synthesis. It is bactericidal for both Gram-positive and Gram-negative organisms and therefore useful for empiric broad-spectrum antibacterial coverage. (Note MRSA resistance to this class.). Toxicity may include gastrointestinal upset and diarrhea, nausea, seizures, headache, rash and allergic reactions. Examples include Ertapenem, Doripenem, Imipenem/Cilastatin, Meropenem.

Cephalosporins (First generation). Have the same mode of action as other beta-lactam antibiotic to disrupt the synthesis of the peptidoglycan layer of bacterial cell walls. The class provides good coverage against Gram positive infections. Potential toxicities include gastrointestinal upset and diarrhea, nausea (if alcohol taken concurrently) and allergic reactions. Examples include Cefadroxil, Cefazolin, Cefalotin, Cefalothin, Keflin, and Cefalexin.

Cephalosporins (Second generation). This class provides less gram-positive coverage than the above with improved gram negative cover. They have the same mode of action as other beta-lactam antibiotics and disrupt the synthesis of the peptidoglycan layer of bacterial cell walls. They may cause gastrointestinal upset and diarrhea, nausea (if alcohol taken concurrently) and allergic reactions. Examples include: Cefaclor, Cefamandole, Cefoxitin, Cefprozil and Cefuroxime.

Cephalosporins (Third generation). Same mode of action as other beta-lactam antibiotic to disrupt the synthesis of the peptidoglycan layer of bacterial cell wall. Provides improved coverage of Gram-negative organisms, except Pseudomonas. Has reduced Gram-positive coverage. May cause gastrointestinal upset and diarrhea, nausea (if alcohol taken concurrently and allergic reactions. Examples include Cefixime, Cefdinir, Cefditoren, Cefoperazone, Cefotaxime, Cefpodoxime, Ceftazidime, Ceftibuten, Ceftizoxime, and Ceftriaxone.

Cephalosporins (Fourth generation). As above for mechanism and toxicity but good coverage for pseudomonal infections. Examples include Cefepime.

Cephalosporins (Fifth generation). As above for mechanism and toxicity but good coverage for Methicillin-resistant Staphylococcus aureus/MRSA. Examples include Ceftaroline fosamil, and Ceftobiprole.

Glycopeptides Inhibit peptidoglycan synthesis and are active against aerobic and anaerobic Gram positive bacteria including MRSA; Vancomycin is used orally for the treatment of C. difficile. Examples include Teicoplanin, Vancomycin, and Telavancin

Lincosamides. Bind to 50S subunit of bacterial ribosomal RNA thereby inhibiting protein synthesis. Used to treat serious staph-, pneumo-, and streptococcal infections in penicillin-allergic patients, also anaerobic infections; clindamycin topically used for acne and possible C. difficile-related pseudomembranous enterocolitis. include Clindamycin and Lincomycin.

Lipopeptides. Bind to the membrane and cause rapid depolarization, resulting in a loss of membrane potential leading to inhibition of protein, DNA and RNA synthesis Gram-positive organisms. Example is Daptomycin.

Macrolides. Are enzyme inhibitors of bacterial protein biosynthesis by binding reversibly to the subunit 50S of the bacterial ribosome, thereby inhibiting translocation of peptidyl-tRNA. Used to treat Streptococcal infections, syphilis, upper respiratory tract infections, lower respiratory tract infection, mycoplasmal infections, Lyme disease. Can cause nausea, vomiting, and diarrhea (especially at higher doses), prolonged QT interval (especially erythromycin) and Jaundice. Examples include Azithromycin, Clarithromycin, irithromycin, Erythromycin, Roxithromycin, Troleandomycin, Telithromycin and Spiramycin.

Monobactams. Same mode of action as other beta-lactam antibiotics, to disrupt the synthesis of the peptidoglycan layer of bacterial cell walls. Example includes Aztreonam.

Nitrofurans. Are used to treat bacterial or protozoal diarrhea or enteritis. An example is Furazolidone and Nitrofurantoin to treat urinary tract infections.

Oxazolidonones. Protein synthesis inhibitors, they prevent the initiation step and are used to treat vancomycin-resistant Staphylococcus aureus. Can cause thrombocytopenia, and peripheral neuropathy. Examples include Linezolid, Radezolid,

Penicillins. Disrupt the synthesis of the peptidoglycan layer of bacterial cell walls.

These are used to treat a wide range of infections; penicillin is used for streptococcal infections, syphilis and Lyme disease and can cause gastrointestinal upset and diarrhea, allergy with serious anaphylactic reaction, brain and kidney damage (rare). Examples include, Amoxicillin, Ampicillin, Azlocillin, Carbenicillin, Cloxacillin, Dicloxacillin, Flucloxacillin, Mezlocillin, Methicillin, Nafcillin, Oxacillin, Penicillin G, Penicillin V, Piperacillin, Penicillin G, Temocillin, Ticarcillin.

Penicillin combinations. The second component prevents bacterial antibiotic resistance to the first component. Examples include Augmentin, Ampicillin/sulbactam, Piperacillin/tazobactam, Ticarcillin/clavulanate.

Polypeptide antibiotics. For treatment of eye, ear or bladder infections; usually applied directly to the eye or inhaled into the lungs; rarely given by injection, although the use of intravenous colistin is experiencing a resurgence due to the emergence of multi drug resistant organisms. This class can cause kidney and nerve damage (when given by injection). The class inhibits isoprenyl pyrophosphate, a molecule that carries the building blocks of the peptidoglycan bacterial cell wall outside of the inner membrane. Examples include Bacitracin, Colistin, and Polymyxin B

Quinolones. For treatment of urinary tract infections, bacterial prostatitis, community-acquired pneumonia, bacterial diarrhea, mycoplasmal infection, gonorrhea. Can cause nausea (rare), irreversible damage to central nervous system (uncommon), tendinosis (rare). The class works by inhibiting the bacterial DNA gyrase or the topoisomerase IV enzyme, thereby inhibiting DNA replication and transcription. Examples include, Ciprofloxacin, Enoxacin, Gatifloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Avelox, Nalidixic acid, Norfloxacin, Ofloxacin, Trovafloxacin, Grepafloxacin, Raxar, Sparfloxacin and Temafloxacin.

Sulfonamides. They are competitive inhibitors of the enzyme dihydropteroate synthetase, DHPS. DHPS catalyses the conversion of PABA (para-Aminobenzoic acid) to dihydropteroic acid|dihydropteroate, a key step in folate synthesis. Folate is necessary for the cell to synthesize nucleic acids (nucleic acids are essential building blocks of DNA and RNA, and in its absence cells will be unable to divide. The class is used to treat Urinary tract infections (except sulfacetamide, used for Conjunctivitis, and mafenide and silver sulfadiazine, used topically for burns. The class can cause nausea, vomiting, and diarrhea, Allergy, including skin rashes, crystals in urine, Renal failure, decrease in white blood cell count and sensitivity to sunlight. Examples include Mafenide, Sulfacetamide, Sulfadiazine, Silver sulfadiazine, Sulfadimethoxine, Sulfamethizole, Sulfamethoxazole, Sulfanilimide, Sulfasalazine, Sulfisoxazole, and Trimethoprim-Sulfamethoxazole.

Tetracyclines Inhibit the binding of aminoacyl-tRNA to the mRNA-ribosome complex. They do so mainly by binding to the 30S ribosomal subunit in the mRNA translation complex. Can be used to treat Syphilis, Chlamydia infections, Lyme disease, mycoplasmal infections, acne, rickettsial infections, and malaria caused by a protest and not a bacterium. Toxicity includes Gastrointestinal upset, Sensitivity to sunlight, Potential toxicity to mother and fetus during pregnancy, Enamel hypoplasia (staining of teeth; potentially permanent, transient depression of bone growth. Examples include Demeclocycline, Doxycycline, Minocycline, Oxytetracycline, and Tetracycline.

Drugs against mycobacteria include the following: Clofazimine, Dapsone, Capreomycin, Cycloserine, Ethambutol, Ethionamide, Isoniazid, Pyrazinamide, Rifampicin, Rifabutin, Rifapentine, Streptomycin, and aminoglycosides.

Other antibiotics include the following:

Arsphenamine, Chloramphenicol, Fosfomycin, Fusidic acid, Metronidazole, Mupirocin, Platensimycin, Quinupristin/Dalfopristin, Thiamphenicol, Tigecycline, Tinidazole, and Trimethoprim. Anti-Viral Medications by Indication

Herpes Simplex Virus (HSV), Varicella Zoster Virus (VZV) and cytomegalovirus (CMV). Oral herpes simplex virus (HSV) causes mucous membrane lesions (i.e., cold sores), and genital HSV causes genital herpetic lesions. Treatment for HSV can also be used for the treatment of Varicella Zoster Virus (VZV) the causative agent for chicken-pox in children and shingles in adults. Typical anti-virals include Acyclovir and Valaciclovir, both inhibitors of viral DNA synthesis. Additionally, Idoxuridine and Brivudin can be incorporated into the viral DNA leading to a hindered mechanism of DNA duplication. A third type of herpes viruses with established treatment is cytomegalovirus (CMV), particularly dangerous for unborn children, infants and immune-compromised patients. Medications used to treat CMV are Ganciclovir and Foscarnet, also indicated in some HSV infections. They act to inhibit viral DNA synthesis.

HIV. A diverse group of antiviral medications control viral load, but cannot cure HIV infections. Viral entry inhibitors such as Enfuvirtide prevent newly formed viruses from entering uninfected host cells by preventing virus-cell fusion.

Reverse transcriptase inhibitors include many drugs such as Abacavir, Lamivudine, Zidovudine, Tenofovir, Efavirenz and Nevirapine. These drugs inhibit reverse transcriptase, an enzyme critical to the mechanism by which HIV transcribes genetic material.

Another anti-viral approach utilizes the protease inhibitors such as Atazanavir, Indinavirn and Ritonavir to inhibit assembly of new viruses. Combination therapies using 2 or 3 of the aforementioned agents are very effective at reducing serum viral load to below detectable levels.

Hepatitis. One of the few anti-HBV (hepatitis B) medications is Lamivudine, a reverse transcriptase inhibitor. Additionally, adefovir and dipivoxil, medications used in the treatment of HIV can be used to inhibit transcription of viral HBV RNA into DNA. Interferons are naturally occurring molecules that stimulate immune responses against invading species, including viral particles. Imiquimod up-regulates the natural production of interferons to boost the human immune response. Synthetically produced Alpha-interferon is also effective in treating HBV and HCV, especially in combination with other drugs. Unfortunately, interferons are associated with a number of severe toxicities that limit their long-terms usage in a number of patients.

Broad-spectrum Antiviral Medications

Ribavirin is effective in the treatment of influenza, HCV and paramyxoviruses such as measles and respiratory syncytial virus by blocking synthesis of viral RNA. A combination of Ribavirin and Alfa-interferon is proven to be effective in treatment of chronic hepatitis C infections.

Inflammation. Anti-Inflammatory medications by class

Glucocorticoids. This class of anti-inflammatory medication reduces inflammation by binding to glucocorticoid receptors (GR). The activated GR complex, in turn, up-regulates the expression of anti-inflammatory proteins in the nucleus (a process known as transactivation) and represses the expression of pro-inflammatory proteins in the cytosol by preventing the translocation of other transcription factors from the cytosol into the nucleus. These drugs are often referred to as corticosteroids. Examples include Budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone and prednisolone.

Non-steroidal anti-inflammatory drugs (NSAIDs). NSAIDs reduce inflammation by reducing the production of prostaglandins, chemicals that promote inflammation, pain, and fever. Prostaglandins also protect the lining of the stomach and intestines from the damaging effects of acid, and promote blood clotting by activating blood platelets and affect kidney function. The enzymes that produce prostaglandins are called cyclooxygenase (COX). There are two types of COX enzymes, COX-1 and COX-2. Both enzymes produce prostaglandins that promote inflammation, pain, and fever; however, only COX-1 produces prostaglandins that activate platelets and protect the stomach and intestinal lining. NSAIDs block COX enzymes and reduce production of prostaglandins. Therefore, inflammation, pain, and fever are reduced. Since the prostaglandins that protect the stomach and promote blood clotting also are reduced, NSAIDs can cause ulcers in the stomach and intestines, and increase the risk of bleeding. Aspirin is the only NSAID that inhibits the clotting of blood for a prolonged period of time, four to seven days, and is therefore effective for preventing blood clots that cause heart attacks and strokes. Ketorolac is a very potent NSAID and is used for treating severe pain that normally would be managed with narcotics. Ketorolac causes ulcers more frequently than other NSAIDs and should not be used for more than five days. Celecoxib blocks COX-2 but has little effect on COX-1. Therefore, celecoxib is sub-classified as a selective COX-2 inhibitor, and it causes fewer ulcers and less bleeding than other NSAIDs. Commonly prescribed NSAIDs include aspirin, salsalate, celecoxib, diclofenac, etodolac, ibuprofen, indomethacin, ketoprofen, ketorolac, nabumetone, naproxen, oxaprozin, piroxicam, sulindac and tolmetin.

Neurological Diseases

Huntington's Disease and dyskinesias. Chorea is an abnormal involuntary movement disorder, one of a group of neurological disorders called dyskinesias, which are caused by overactivity of the neurotransmitter dopamine in the areas of the brain that control movement. Chorea is characterized by brief, irregular contractions that are not repetitive or rhythmic, but appear to flow from one muscle to the next. Chorea often occurs with athetosis, which adds twisting and writhing movements. Chorea is a primary feature of Huntington's disease, a progressive, hereditary movement disorder that appears in adults, but it may also occur in a variety of other conditions. Syndenham's chorea occurs in a small percentage (20 percent) of children and adolescents as a complication of rheumatic fever. Chorea can also be induced by drugs (levodopa, anti-convulsants, and anti-psychotics) metabolic and endocrine disorders, and vascular incidents. There is currently no standard course of treatment for chorea. Treatment depends on the type of chorea and the associated disease. Treatment for Huntington's disease is supportive, while treatment for Syndenham's chorea usually involves antibiotic drugs to treat the infection, followed by drug therapy to prevent recurrence. Adjusting medication dosages can treat drug-induced chorea. Metabolic and endocrine-related choreas are treated according to the cause(s) of symptoms.

Parkinson's Disease. Parkinson's disease (PD) belongs to a group of conditions called motor system disorders, which are the result of the loss of dopamine-producing brain cells. The four primary symptoms of PD are tremor, or trembling in hands, arms, legs, jaw, and face; rigidity, or stiffness of the limbs and trunk; bradykinesia, or slowness of movement; and postural instability, or impaired balance and coordination. PD usually affects people over the age of 50. Other symptoms may include depression and other emotional changes; difficulty in swallowing, chewing, and speaking; urinary problems or constipation; skin problems; and sleep disruptions. There are currently no blood or laboratory tests that have been proven to help in diagnosing sporadic PD. Therefore the diagnosis is based on medical history and a neurological examination. The disease can be difficult to diagnose accurately. There is no cure for PD, but a variety of medications are used to relieve symptoms. Patients are given levodopa combined with carbidopa. Carbidopa delays the conversion of levodopa into dopamine until it reaches the brain. Nerve cells can use levodopa to make dopamine and replenish the brain supply. Anticholinergics may help control tremor and rigidity. Other drugs, such as bromocriptine, pramipexole, and ropinirole, mimic the role of dopamine in the brain, causing the neurons to react as they would to dopamine. An antiviral drug, amantadine, also appears to reduce symptoms. Rasagiline can be used along with levodopa for patients with advanced PD or as a single-drug treatment for early PD. In some cases, surgery may be appropriate if the disease doesn't respond to drugs. A therapy called deep brain stimulation (DBS) has now been approved by the U.S. Food and Drug Administration. In DBS, electrodes are implanted into the brain and connected to a small electrical device called a pulse generator that can be externally programmed. DBS can reduce the need for levodopa and related drugs, which in turn decreases the involuntary movements called dyskinesias that are a common side effect of levodopa. It also helps to alleviate fluctuations of symptoms and to reduce tremors, slowness of movements, and gait problems. DBS requires careful programming of the stimulator device in order to work correctly.

Amyotrophic Lateral Sclerosis. Amyotrophic lateral sclerosis (ALS), sometimes called Lou Gehrig's disease or classical motor neuron disease, is a rapidly progressive, invariably fatal neurological disease that attacks the neurons responsible for controlling voluntary muscles. In ALS, both the upper motor neurons and the lower motor neurons degenerate or die, ceasing to send messages to muscles. Unable to function, the muscles gradually atrophy. Symptoms are usually first noticed in the arms and hands, legs, or swallowing muscles. Muscle weakness and atrophy occur on both sides of the body. Individuals with ALS lose their strength and the ability to move their arms and legs, and to hold the body upright. The disease does not affect a person's ability to see, smell, taste, hear, or recognize touch. Although the disease does not usually impair a person's mind or personality, several recent studies suggest that some people with ALS may develop cognitive problems involving word fluency, decision-making, and memory. The cause of ALS is not known. No cure has yet been found for ALS. The drug riluzole prolongs life by 2-3 months but does not relieve symptoms.

Multiple Sclerosis. Multiple sclerosis (MS) is a neurologic disease that can range from benign to completely disabling. MS results from an auto-immune response to nerve-insulating myelin. Such assaults may be linked to an unknown environmental trigger, perhaps a virus.

Most people experience their first symptoms of MS between the ages of 20 and 40; the initial symptom of MS is often blurred or double vision, red-green color distortion, or even blindness in one eye. Most MS patients experience muscle weakness in their extremities and difficulty with coordination and balance. These symptoms may be severe enough to impair walking or even standing. In the worst cases, MS can produce partial or complete paralysis. Most people with MS also exhibit paresthesias, transitory abnormal sensory feelings such as numbness, prickling, or “pins and needles” sensations. Some may also experience pain. Speech impediments, tremors, and dizziness are other frequent complaints. Occasionally, people with MS have hearing loss. Approximately half of all people with MS experience cognitive impairments such as difficulties with concentration, attention, memory, and poor judgment, but such symptoms are usually mild and are frequently overlooked. Depression is another common feature of MS. There is as yet no cure for MS. Three forms of beta interferon (Avonex, Betaseron, and Rebif) have now been approved by the Food and Drug Administration for treatment of relapsing-remitting MS. Beta interferon has been shown to reduce the number of exacerbations and may slow the progression of physical disability. When attacks do occur, they tend to be shorter and less severe. The FDA also has approved a synthetic form of myelin basic protein, called copolymer I (Copaxone), for the treatment of relapsing-remitting MS. An immunosuppressant treatment, Novantrone (mitoxantrone), is approved by the FDA for the treatment of advanced or chronic MS. The FDA has also approved dalfampridine (Ampyra) to improve walking in individuals with MS. While steroids do not affect the course of MS over time, they can reduce the duration and severity of attacks in some patients. Spasticity, which can occur either as a sustained stiffness caused by increased muscle tone or as spasms that come and go, is usually treated with muscle relaxants and tranquilizers such as baclofen, tizanidine, diazepam, clonazepam, and dantrolene. Other drugs that may reduce fatigue in some, but not all, patients include amantadine (Symmetrel), pemoline (Cylert), and the still-experimental drug aminopyridine. Although improvement of optic symptoms usually occurs even without treatment, a short course of treatment with intravenous methylprednisolone (Solu-Medrol) followed by treatment with oral steroids is sometimes used.

Alzheimer's Disease. Alzheimer's disease is an irreversible, progressive brain disease that slowly destroys memory and thinking skills. In most people with Alzheimer's, symptoms first appear after age 60. Estimates vary, but as many as 5.1 million Americans may have Alzheimer's disease. Patient's exhibit various brain abnormalities including amyloid plaques, neurofibrillary tangles, and neuronal loss. Four medications are approved by the U.S. Food and Drug Administration to treat Alzheimer's. Donepezil, rivastigmine and galantamine are used to treat mild to moderate Alzheimer's. Memantine is used to treat moderate to severe Alzheimer's. These drugs do not change the underlying disease process, are effective for some but not all people, and may help only for a limited time.

Schizophrenia. Schizophrenics display three broad categories of symptoms characterized as positive, negative and cognitive. Positive symptoms are psychotic behaviors including hallucinations, delusions, thought and movement disorders. Negative symptoms are associated with disruptions to normal behaviors. These symptoms include flat affect, lack of pleasure in everyday activities, lack of ability to begin and sustain planned activities, and speaking little, even when forced to interact as well as having neglect for basic personal hygiene. Cognitive symptoms include poor ability to understand information and use it to make decisions, trouble focusing or paying attention and problems with the ability to use information immediately after learning it. This neurologic disorder effects 1 percent of the general population, but it occurs in 10 percent of people who have a first-degree relative with the disorder. The risk is highest for an identical twin of a person with schizophrenia with a 40-65 percent chance of developing the disorder. No gene causes the disease by itself. Aberrant dopamine and glutamate transmission is believed to play a role in schizophrenia. Treatments include antipsychotic medications and various psychosocial treatments. Older antipsychotic medications include Chlorpromazine, Haloperidol, Perphenazine, Etrafon and Fluphenazine. New antipsychotic medications include clozapine which can cause agranulocytosis, requiring bi-weekly WBC count evaluation. Other atypical antipsychotics include Risperidone, Olanzapine, Quetiapine, Ziprasidone, Aripiprazole and Paliperidone. Side effects of many antipsychotics include drowsiness, dizziness when changing positions, blurred vision, rapid heartbeat, sensitivity to the sun, Skin rashes and menstrual problems for women. Atypical antipsychotic medications can cause major weight gain and changes in a person's metabolism. This may increase a person's risk of getting diabetes and high cholesterol. Typical antipsychotic medications can cause side effects related to physical movement, such as rigidity, persistent muscle spasms, tremors and restlessness. Long-term use of typical antipsychotic medications may lead to a condition called tardive dyskinesia (TD). TD causes uncontrolled, and in some cases permanent, involuntary muscle movements.

Additional Description of the Invention

The present invention relates to methods and compositions for the treatment of any gene that is desirable to modulate expression of. This includes but is not limited to cancers. In the next sections will will describe both cancer and non-cancer targets and then in the section immediately following those selected cancer and non-cancer targets we will present over 40 High Value Targets, both cancer and noncancer, with sequence information, and some of these examples will have data with detailed information about our techniques and methods as well as our surprising results.

Cancer Targets

In some embodiments, the present invention provides oligonucleotide-based therapeutics for the inhibition of oncogenes involved in a variety of cancers. The present invention is not limited to the treatment of cancer or any particular cancer. Any cancer can be targeted, including, but not limited to, breast cancers. The present invention is also not limited to the targeting of cancers or oncogenes. The methods and compositions of the present invention are suitable for use with any gene that it is desirable to inhibit the expression of (e.g., for therapeutic or research uses. Specific gene targets that have been optimally identified as susceptible to the DNAi therapeutic approach are described below.

Oncogene Targets such as,

In some embodiments, the present invention provides DNAi inhibitors of oncogenes. The present invention is not limited to the inhibition of a particular oncogene. Indeed, the present invention encompasses DNAi inhibitors to any number of oncogenes including, but not limited to, those disclosed herein.

Combination Therapies with Cancer Targets

In some embodiments, the compositions of the present invention are provided in combination with existing therapies. In other embodiments, two or more compounds of the present invention are provided in combination. In some embodiments, the compounds of the present invention are provided in combination with known cancer chemotherapy agents. The present invention is not limited to a particular chemotherapy agent.

Various classes of antineoplastic (e.g., anticancer) agents are contemplated for use in certain embodiments of the present invention. Anticancer agents suitable for use with the present invention include, but are not limited to, agents that induce apoptosis, agents that inhibit adenosine deaminase function, inhibit pyrimidine biosynthesis, inhibit purine ring biosynthesis, inhibit nucleotide interconversions, inhibit ribonucleotide reductase, inhibit thymidine monophosphate (TMP) synthesis, inhibit dihydrofolate reduction, inhibit DNA synthesis, form adducts with DNA, damage DNA, inhibit DNA repair, intercalate with DNA, deaminate asparagines, inhibit RNA synthesis, inhibit protein synthesis or stability, inhibit microtubule synthesis or function, and the like.

In some embodiments, exemplary anticancer agents suitable for use in compositions and methods of the present invention include, but are not limited to: 1) alkaloids, including microtubule inhibitors (e.g., vincristine, vinblastine, and vindesine, etc.), microtubule stabilizers (e.g., paclitaxel (TAXOL), and docetaxel, etc.), and chromatin function inhibitors, including topoisomerase inhibitors, such as epipodophyllotoxins (e.g., etoposide (VP-16), and teniposide (VM-26), etc.), and agents that target topoisomerase I (e.g., camptothecin and isirinotecan (CPT-11), etc.); 2) covalent DNA-binding agents (alkylating agents), including nitrogen mustards (e.g., mechlorethamine, chlorambucil, cyclophosphamide, ifosphamide, and busulfan (MYLERAN), etc.), nitrosoureas (e.g., carmustine, lomustine, and semustine, etc.), and other alkylating agents (e.g., dacarbazine, hydroxymethylmelamine, thiotepa, and mitomycin, etc.); 3) noncovalent DNA-binding agents (antitumor antibiotics), including nucleic acid inhibitors (e.g., dactinomycin (actinomycin D), etc.), anthracyclines (e.g., daunorubicin (daunomycin, and cerubidine), doxorubicin (adriamycin), and idarubicin (idamycin), etc.), anthracenediones (e.g., anthracycline analogues, such as mitoxantrone, etc.), bleomycins (BLENOXANE), etc., and plicamycin (mithramycin), etc.; 4) antimetabolites, including antifolates (e.g., methotrexate, FOLEX, and MEXATE, etc.), purine antimetabolites (e.g., 6-mercaptopurine (6-MP, PURINETHOL), 6-thioguanine (6-TG), azathioprine, acyclovir, ganciclovir, chlorodeoxyadenosine, 2-chlorodeoxyadenosine (CdA), and 2′-deoxycoformycin (pentostatin), etc.), pyrimidine antagonists (e.g., fluoropyrimidines (e.g., 5-fluorouracil (ADRUCIL), 5-fluorodeoxyuridine (FdUrd) (floxuridine)) etc.), and cytosine arabinosides (e.g., CYTOSAR (ara-C) and fludarabine, etc.); 5) enzymes, including L-asparaginase, and hydroxyurea, etc.; 6) hormones, including glucocorticoids, antiestrogens (e.g., tamoxifen, etc.), nonsteroidal antiandrogens (e.g., flutamide, etc.), and aromatase inhibitors (e.g., anastrozole (ARIMIDEX), etc.); 7) platinum compounds (e.g., cisplatin and carboplatin, etc.); 8) monoclonal antibodies conjugated with anticancer drugs, toxins, and/or radionuclides, etc.; 9) biological response modifiers (e.g., interferons (e.g., IFN-α, etc.) and interleukins (e.g., IL-2, etc.), etc.); 10) adoptive immunotherapy; 11) hematopoietic growth factors; 12) agents that induce tumor cell differentiation (e.g., all-trans-retinoic acid, etc.); 13) gene therapy techniques; 14) antisense therapy techniques; 15) tumor vaccines; 16) therapies directed against tumor metastases (e.g., batimastat, etc.); 17) angiogenesis inhibitors; 18) proteosome inhibitors (e.g., VELCADE); 19) inhibitors of acetylation and/or methylation (e.g., HDAC inhibitors); 20) modulators of NF kappa B; 21) inhibitors of cell cycle regulation (e.g., CDK inhibitors); 22) modulators of p53 protein function; and 23) radiation.

Any oncolytic agent that is routinely used in a cancer therapy context finds use in the compositions and methods of the present invention. For example, the U.S. Food and Drug Administration maintains a formulary of oncolytic agents approved for use in the United States. International counterpart agencies to the U.S.F.D.A. maintain similar formularies. Table 1 provides a list of exemplary antineoplastic agents approved for use in the U.S. Those skilled in the art will appreciate that the “product labels” required on all U.S. approved chemotherapeutics describe approved indications, dosing information, toxicity data, and the like, for the exemplary agents.

TABLE 1 Aldesleukin Proleukin Chiron Corp., (des-alanyl-1, serine-125 human interleukin-2) Emeryville, CA Alemtuzumab Campath Millennium and (IgG1κ anti CD52 antibody) ILEX Partners, LP, Cambridge, MA Alitretinoin Panretin Ligand (9-cis-retinoic acid) Pharmaceuticals, Inc., San Diego CA Allopurinol Zyloprim GlaxoSmithKline, (1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one Research Triangle monosodium salt) Park, NC Altretamine Hexalen US Bioscience, (N,N,N′,N′,N″,N″,-hexamethyl-1,3,5-triazine-2,4,6- West triamine) Conshohocken, PA Amifostine Ethyol US Bioscience (ethanethiol, 2-[(3-aminopropyl)amino]-, dihydrogen phosphate (ester)) Anastrozole Arimidex AstraZeneca (1,3-Benzenediacetonitrile,a,a,a′,a′-tetramethyl-5- Pharmaceuticals, (1H-1,2,4-triazol-1-ylmethyl)) LP, Wilmington, DE Arsenic trioxide Trisenox Cell Therapeutic, Inc., Seattle, WA Asparaginase Elspar Merck & Co., Inc., (L-asparagine amidohydrolase, type EC-2) Whitehouse Station, NJ BCG Live TICE BCG Organon Teknika, (lyophilized preparation of an attenuated strain of Corp., Durham, NC Mycobacterium bovis (Bacillus Calmette-Gukin [BCG], substrain Montreal) bexarotene capsules Targretin Ligand (4-[1-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2- Pharmaceuticals napthalenyl) ethenyl] benzoic acid) Bexarotene gel Targretin Ligand Pharmaceuticals Bleomycin Blenoxane Bristol-Myers (cytotoxic glycopeptide antibiotics produced by Squibb Co., NY, Streptomyces verticillus; bleomycin A2 and NY bleomycin B2) Capecitabine Xeloda Roche (5′-deoxy-5-fluoro-N-[(pentyloxy)carbonyl]-cytidine) Carboplatin Paraplatin Bristol-Myers (platinum, diammine [1,1- Squibb cyclobutanedicarboxylato(2-)-0,0′]-,(SP-4-2)) Carmustine BCNU, Bristol-Myers (1,3-bis(2-chloroethyl)-1-nitrosourea) BiCNU Squibb Carmustine with Polifeprosan 20 Implant Gliadel Guilford Wafer Pharmaceuticals, Inc., Baltimore, MD Celecoxib Celebrex Searle (as 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H- Pharmaceuticals, pyrazol-1-yl] benzenesulfonamide) England Chlorambucil Leukeran GlaxoSmithKline (4-[bis(2chlorethyl)amino]benzenebutanoic acid) Cisplatin Platinol Bristol-Myers (PtC12H6N2) Squibb Cladribine Leustatin, 2- R. W. Johnson (2-chloro-2′-deoxy-b-D-adenosine) CdA Pharmaceutical Research Institute, NJ Cyclophosphamide Cytoxan, Bristol-Myers (2-[bis(2-chloroethyl)amino] tetrahydro-2H-13,2- Neosar Squibb oxazaphosphorine 2-oxide monohydrate) Cytarabine Cytosar-U Pharmacia & (1-b-D-Arabinofuranosylcytosine, C9H13N3O5) Upjohn Company Cytarabine liposomal DepoCyt Skye Pharmaceuticals, Inc., San Diego, CA Dacarbazine DTIC-Dome Bayer AG, (5-(3,3-dimethyl-1-triazeno)-imidazole-4- Leverkusen, carboxamide (DTIC)) Germany Dactinomycin, actinomycin D Cosmegen Merck (actinomycin produced by Streptomyces parvullus, C62H86N12O16) Darbepoetin alfa Aranesp Amgen, Inc., (recombinant peptide) Thousand Oaks, CA daunorubicin liposomal DanuoXome Nexstar ((8S-cis)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-á-L- Pharmaceuticals, lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11- Inc., Boulder, CO trihydroxy-1-methoxy-5,12-naphthacenedione hydrochloride) Daunorubicin HCl, daunomycin Cerubidine Wyeth Ayerst, ((1S,3S)-3-Acetyl-1,2,3,4,6,11-hexahydro-3,5,12- Madison, NJ trihydroxy-10-methoxy-6,11-dioxo-1-naphthacenyl 3-amino-2,3,6-trideoxy-(alpha)-L-lyxo- hexopyranoside hydrochloride) Denileukin diftitox Ontak Seragen, Inc., (recombinant peptide) Hopkinton, MA Dexrazoxane Zinecard Pharmacia & ((S)-4,4′-(1-methyl-1,2-ethanediyl)bis-2,6- Upjohn Company piperazinedione) Docetaxel Taxotere Aventis ((2R,3S)-N-carboxy-3-phenylisoserine, N-tert-butyl Pharmaceuticals, ester, 13-ester with 5b-20-epoxy-12a,4,7b,10b,13a- Inc., Bridgewater, hexahydroxytax-11-en-9-one 4-acetate 2-benzoate, NJ trihydrate) Doxorubicin HCl Adriamycin, Pharmacia & (8S,10S)-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo- Rubex Upjohn Company hexopyranosyl)oxy]-8-glycolyl-7,8,9,10-tetrahydro- 6,8,11-trihydroxy-1-methoxy-5,12- naphthacenedione hydrochloride) doxorubicin Adriamycin Pharmacia & PFS Upjohn Company Intravenous injection doxorubicin liposomal Doxil Sequus Pharmaceuticals, Inc., Menlo park, CA dromostanolone propionate Dromostanolone Eli Lilly & (17b-Hydroxy-2a-methyl-5a-androstan-3-one Company, propionate) Indianapolis, IN dromostanolone propionate Masterone Syntex, Corp., Palo injection Alto, CA Elliott′s B Solution Elliott′s B Orphan Medical, Solution Inc Epirubicin Ellence Pharmacia & ((8S-cis)-10-[(3-amino-2,3,6-trideoxy-a-L-arabino- Upjohn Company hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11- trihydroxy-8-(hydroxyacetyl)-1-methoxy-5,12- naphthacenedione hydrochloride) Epoetin alfa Epogen Amgen, Inc (recombinant peptide) Estramustine Emcyt Pharmacia & (estra-1,3,5(10)-triene-3,17-diol(17(beta))-, 3-[bis(2- Upjohn Company chloroethyl)carbamate] 17-(dihydrogen phosphate), disodium salt, monohydrate, or estradiol 3-[bis(2- chloroethyl)carbamate] 17-(dihydrogen phosphate), disodium salt, monohydrate) Etoposide phosphate Etopophos Bristol-Myers (4′-Demethylepipodophyllotoxin9-[4,6-O-(R)- Squibb ethylidene-(beta)-D-glucopyranoside], 4′- (dihydrogen phosphate)) etoposide, VP-16 Vepesid Bristol-Myers (4′-demethylepipodophyllotoxin 9-[4,6-0-(R)- Squibb ethylidene-(beta)-D-glucopyranoside]) Exemestane Aromasin Pharmacia & (6-methylenandrosta-1,4-diene-3,17-dione) Upjohn Company Filgrastim Neupogen Amgen, Inc (r-metHuG-CSF) floxuridine (intraarterial) FUDR Roche (2′-deoxy-5-fluorouridine) Fludarabine Fludara Berlex (fluorinated nucleotide analog of the antiviral agent Laboratories, Inc., vidarabine, 9-b-D-arabinofuranosyladenine (ara-A)) Cedar Knolls, NJ Fluorouracil, 5-FU Adrucil ICN (5-fluoro-2,4(1H,3H)-pyrimidinedione) Pharmaceuticals, Inc., Humacao, Puerto Rico Fulvestrant Faslodex IPR (7-alpha-[9-(4,4,5,5,5-penta fluoropentylsulphinyl) Pharmaceuticals, nonyl]estra-1,3,5-(10)-triene-3,17-beta-diol) Guayama, Puerto Rico Gemcitabine Gemzar Eli Lilly (2′-deoxy-2′,2′-difluorocytidine monohydrochloride (b-isomer)) Gemtuzumab Ozogamicin Mylotarg Wyeth Ayerst (anti-CD33 hP67.6) Goserelin acetate Zoladex AstraZeneca (acetate salt of [D-Ser(But)6,Azgly10]LHRH; pyro- Implant Pharmaceuticals Glu-His-Trp-Ser-Tyr-D-Ser(But)-Leu-Arg-Pro- Azgly-NH2 acetate [C59H84N18O14•(C2H4O2)x Hydroxyurea Hydrea Bristol-Myers Squibb Ibritumomab Tiuxetan Zevalin Biogen IDEC, Inc., (immunoconjugate resulting from a thiourea covalent Cambridge MA bond between the monoclonal antibody Ibritumomab and the linker-chelator tiuxetan [N-[2- bis(carboxymethyl)amino]-3-(p- isothiocyanatophenyl)-propyl]-[N-[2- bis(carboxymethyl)amino]-2-(methyl)- ethyl]glycine) Idarubicin Idamycin Pharmacia & (5,12-Naphthacenedione, 9-acetyl-7-[(3-amino- Upjohn Company 2,3,6-trideoxy-(alpha)-L-lyxo-hexopyranosyl)oxy]- 7,8,9,10-tetrahydro-6,9,11-trihydroxyhydrochloride, (7S-cis)) Ifosfamide IFEX Bristol-Myers (3-(2-chloroethyl)-2-[(2- Squibb chloroethyl)amino]tetrahydro-2H-1,3,2- oxazaphosphorine 2-oxide) Imatinib Mesilate Gleevec Novartis AG, (4-[(4-Methyl-1-piperazinyl)methyl]-N-[4-methyl-3- Basel, Switzerland [[4-(3 -pyridinyl)-2-pyrimidinyl] amino]- phenyl]benzamide methanesulfonate) Interferon alfa-2a Roferon-A Hoffmann-La (recombinant peptide) Roche, Inc., Nutley, NJ Interferon alfa-2b Intron A Schering AG, (recombinant peptide) (Lyophilized Berlin, Germany Betaseron) Irinotecan HCl Camptosar Pharmacia & ((4S)-4,11-diethyl-4-hydroxy-9-[(4-piperi- Upjohn Company dinopiperidino)carbonyloxy]-1H-pyrano[3′,4′:6,7] indolizino[1,2-b] quinoline-3,14(4H,12H) dione hydrochloride trihydrate) Letrozole Femara Novartis (4,4′-(1H-1,2,4-Triazol-1-ylmethylene) dibenzonitrile) Leucovorin Wellcovorin, Immunex, Corp., (L-Glutamic acid, N[4[[(2amino-5-formyl-1,4,5,6,7,8 Leucovorin Seattle, WA hexahydro4oxo6-pteridinyl)methyl] amino]benzoyl] , calcium salt (1:1)) Levamisole HCl Ergamisol Janssen Research ((−)-(S)-2,3,5,6-tetrahydro-6-phenylimidazo [2,1-b] Foundation, thiazole monohydrochloride C11H12N2S•HCl) Titusville, NJ Lomustine CeeNU Bristol-Myers (1-(2-chloro-ethyl)-3-cyclohexyl-1-nitrosourea) Squibb Meclorethamine, nitrogen mustard Mustargen Merck (2-chloro-N-(2-chloroethyl)-N-methylethanamine hydrochloride) Megestrol acetate Megace Bristol-Myers 17α(acetyloxy)-6-methylpregna-4,6-diene-3,20- Squibb dione Melphalan, L-PAM Alkeran GlaxoSmithKline (4-[bis(2-chloroethyl) amino]-L-phenylalanine) Mercaptopurine, 6-MP Purinethol GlaxoSmithKline (1,7-dihydro-6H-purine-6-thione monohydrate) Mesna Mesnex Asta Medica (sodium 2-mercaptoethane sulfonate) Methotrexate Methotrexate Lederle (N-[4-[[(2,4-diamino-6- Laboratories pteridinyl)methyl]methylamino]benzoyl]-L-glutamic acid) Methoxsalen (9-methoxy-7H-furo[3,2-g][1]- Uvadex Therakos, Inc., benzopyran-7-one) Way Exton, Pa Mitomycin C Mutamycin Bristol-Myers Squibb Mitomycin C Mitozytrex SuperGen, Inc., Dublin, CA Mitotane Lysodren Bristol-Myers (1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) Squibb ethane) Mitoxantrone Novantrone Immunex (1,4-dihydroxy-5,8-bis[[2-[(2- Corporation hydroxyethyl)amino]ethyl]amino]-9,10- anthracenedione dihydrochloride) Nandrolone phenpropionate Durabolin-50 Organon, Inc., West Orange, NJ Nofetumomab Verluma Boehringer Ingelheim Pharma KG, Germany Oprelvekin Neumega Genetics Institute, (IL-11) Inc., Alexandria, VA Oxaliplatin Eloxatin Sanofi Synthelabo, (cis-[(1R,2R)-1,2-cyclohexanediamine-N,N′] Inc., NY, NY [oxalato(2-)-O,O′] platinum) Paclitaxel TAXOL Bristol-Myers (5β,20-Epoxy-1,2a,4,7β,10β,13a-hexahydroxytax- Squibb 11-en-9-one 4,10-diacetate 2-benzoate 13-ester with (2R,3S)-N-benzoyl-3-phenylisoserine) Pamidronate Aredia Novartis (phosphonic acid (3-amino-1-hydroxypropylidene) bis-, disodium salt, pentahydrate, (APD)) Pegademase Adagen Enzon ((monomethoxypolyethylene glycol succinimidyl) 11- (Pegademase Pharmaceuticals, 17-adenosine deaminase) Bovine) Inc., Bridgewater, NJ Pegaspargase Oncaspar Enzon (monomethoxypolyethylene glycol succinimidyl L- asparaginase) Pegfilgrastim Neulasta Amgen, Inc (covalent conjugate of recombinant methionyl human G-CSF (Filgrastim) and monomethoxypolyethylene glycol) Pentostatin Nipent Parke-Davis Pharmaceutical Co., Rockville, MD Pipobroman Vercyte Abbott Laboratories, Abbott Park, IL Plicamycin, Mithramycin Mithracin Pfizer, Inc., NY, (antibiotic produced by Streptomyces plicatus) NY Porfimer sodium Photofrin QLT Phototherapeutics, Inc., Vancouver, Canada Procarbazine Matulane Sigma Tau (N-isopropyl-μ-(2-methylhydrazino)-p-toluamide Pharmaceuticals, monohydrochloride) Inc., Gaithersburg, MD Quinacrine Atabrine Abbott Labs (6-chloro-9-(1-methyl-4-diethyl-amine) butylamino-2-methoxyacridine) Rasburicase Elitek Sanofi-Synthelabo, (recombinant peptide) Inc., Rituximab Rituxan Genentech, Inc., (recombinant anti-CD20 antibody) South San Francisco, CA Sargramostim Prokine Immunex Corp (recombinant peptide) Streptozocin Zanosar Pharmacia & (streptozocin 2-deoxy-2- Upjohn Company [[(methylnitrosoamino)carbonyl]amino]-a(and b)- D-glucopyranose and 220 mg citric acid anhydrous) Talc Sclerosol Bryan, Corp., (Mg3Si4O10 (OH)2) Woburn, MA Tamoxifen Nolvadex AstraZeneca ((Z)2-[4-(1,2-diphenyl-1-butenyl) phenoxy]-N,N- Pharmaceuticals dimethylethanamine 2-hydroxy-1,2,3- propanetricarboxylate (1:1)) Temozolomide Temodar Schering (3,4-dihydro-3-methyl-4-oxoimidazo[5,1-d]-as- tetrazine-8-carboxamide) Teniposide, VM-26 Vumon Bristol-Myers (4′-demethylepipodophyllotoxin 9-[4,6-0-(R)-2- Squibb thenylidene-(beta)-D-glucopyranoside]) Testolactone Teslac Bristol-Myers (13-hydroxy-3-oxo-13,17-secoandrosta-1,4-dien-17- Squibb oic acid [dgr]-lactone) Thioguanine, 6-TG Thioguanine GlaxoSmithKline (2-amino-1,7-dihydro-6H-purine-6-thione) Thiotepa Thioplex Immunex (Aziridine,1,1′,1″-phosphinothioylidynetris-, or Tris Corporation (1-aziridinyl) phosphine sulfide) Topotecan HCl Hycamtin GlaxoSmithKline ((S)-10-[(dimethylamino)methyl]-4-ethyl-4,9- dihydroxy-1H-pyrano[3′,4′:6,7] indolizino [1,2-b] quinoline-3,14-(4H,12H)-dione monohydrochloride) Toremifene Fareston Roberts (2-(p-[(Z)-4-chloro-1,2-diphenyl-1-butenyl]- Pharmaceutical phenoxy)-N,N-dimethylethylamine citrate (1:1)) Corp., Eatontown, NJ Tositumomab, I 131 Tositumomab Bexxar Corixa Corp., (recombinant murine immunotherapeutic monoclonal Seattle, WA IgG2a lambda anti-CD20 antibody (I 131 is a radioimmunotherapeutic antibody)) Trastuzumab Herceptin Genentech, Inc (recombinant monoclonal IgG1 kappa anti-HER2 antibody) Tretinoin, ATRA Vesanoid Roche (all-trans retinoic acid) Uracil Mustard Uracil Roberts Labs Mustard Capsules Valrubicin, N-trifluoroacetyladriamycin-14- Valstar Anthra --> Medeva valerate ((2S-cis)-2-[1,2,3,4,6,11-hexahydro-2,5,12- trihydroxy-7 methoxy-6,11-dioxo-[[4 2,3,6-trideoxy- 3-[(trifluoroacetyl)-amino-α-L-lyxo- hexopyranosyl]oxyl]-2-naphthacenyl]-2-oxoethyl pentanoate) Vinblastine, Leurocristine Velban Eli Lilly (C46H56N4O10•H2SO4) Vincristine Oncovin Eli Lilly (C46H56N4O10•H2SO4) Vinorelbine Navelbine GlaxoSmithKline (3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine [R-(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)]) Zoledronate, Zoledronic acid Zometa Novartis ((1-Hydroxy-2-imidazol-1-yl-phosphonoethyl) phosphonic acid monohydrate)

Other identified cancer combination therapies include the following: PI3K inhibitors (CAL101), Bruton Kinase inhibitor (PCI-32765), and BCL-6 inhibitor. This document describes the targets and associated therapy for these identified cancers as being particularly susceptible to treatment with combination therapies. Targets

The present invention is not limited to the cancer and non-cancer targets listed above commonly found in humans. The present invention can also be applied both to other cancer targets (also referred to as oncogenes) (and where such cancer targets may also be involved in other disease such as inflammation, neurological, metabolic, cardiovascular, etc.) and to non-cancer target such as Cardiovascular/Metabolic Disease, Eye Disease, Infectious Disease, Inflammation, Neurological Disease, Rare Disease, and Stem Cells. Examples of specific genes are included in Table 2, but are not limited to those described in Table. Additional targets are not listed but can be found in the key proliferation pathways such as MAPK, PI3K, MEK, etc. The present invention can also apply to disease and growth targets for plant genome and animal genomes.

TABLE 2 Cancer and non-cancer targets DNAi Disease, Gene, and Cell System Targets ID Disease Area Target 1 Cancer 2-dG 2 Cancer 4-1BB 3 Cancer ABCB1 4 Cancer ABL 5 Cancer ABL1/BCR 6 Cancer Act-1 7 Cancer ADAM12 8 Cancer ADAM7 9 Cancer ADAMTS4 10 Cancer ADAMTS5 11 Cancer AFP (Alpha-fetoprotein) 12 Cancer AKT 13 Cancer AKT1 14 Cancer AKT2 15 Cancer AKT3 16 Cancer AldoA 17 Cancer ALK 18 Cancer ALK/NPM1 19 Cancer AMI1 20 Cancer AML1/ETO 21 Cancer Androgen Receptor (AR) 22 Cancer Angiopoeitin (ANG) 23 Cancer ANGPT2 (ANG-2) 24 Cancer APC 25 Cancer ARAF 26 Cancer AR 27 Cancer AREG 28 Cancer ARF6 29 Cancer ARNT 30 Cancer Aromatase Inhibitors (Ais) 31 Cancer ASXL1 32 Cancer ATM 33 Cancer ATRX 34 Cancer AXIN1 35 Cancer AXL 36 Cancer B7H3 37 Cancer BAX 38 Cancer BBC3 39 Cancer BCBL 40 Cancer BCL1 41 Cancer BCL2 42 Cancer BCL2L1 (BCLXL) 43 Cancer BCL2L 11 44 Cancer BCL3 45 Cancer BCL6 46 Cancer BCR/ABL 47 Cancer BDNF 48 Cancer Beclin-1 49 Cancer Beta catenin 50 Cancer BIRC2 (c-IAP1) 51 Cancer BIRC3 (c-IAP2) 52 Cancer BIRC4 53 Cancer BIRC5 54 Cancer BMI1 55 Cancer BMP10 56 Cancer BRAF 57 Cancer BRCA1 58 Cancer BRCA2 59 Cancer BRD3 60 Cancer BTK 61 Cancer BTLA 62 Cancer C/EBPalpha 63 Cancer C5B-9 64 Cancer CANT1 65 Cancer CASP2 66 Cancer CASP3 67 Cancer CASP8 68 Cancer CBFA2T3 69 Cancer CBFB 70 Cancer CBL 71 Cancer CBLB 72 Cancer CBLC 73 Cancer CCND1 74 Cancer CCND3 75 Cancer CCKBR 76 Cancer CCNA1 77 Cancer CCNB1 78 Cancer CD133 79 Cancer CD19 80 Cancer CD20 81 Cancer CD24 82 Cancer CD30 83 Cancer CD33 84 Cancer CD37 85 Cancer CD38 86 Cancer CD4 87 Cancer CD-40 88 Cancer CD40LG 89 Cancer CD44 90 Cancer CD-52 91 Cancer CD74 92 Cancer CD80 93 Cancer CDC42 94 Cancer CDC25A 95 Cancer CDC25B 96 Cancer CDK2 97 Cancer CDK4 98 Cancer CDK4 99 Cancer CDK6 100 Cancer CDK7 101 Cancer CDKN1A 102 Cancer CDKN1C 103 Cancer CDKN2A 104 Cancer CDKN2B 105 Cancer CDKN2C 106 Cancer c-fos 107 Cancer CHEK1 108 Cancer CHEK2 109 Cancer CHMP5 110 Cancer c-ki-RAS 111 Cancer CKIT 112 Cancer CLTC 113 Cancer Clusterin 114 Cancer CMET 115 Cancer COL6A3 116 Cancer CPK 117 Cancer CRAF 118 Cancer CRB 119 Cancer CRBN 120 Cancer CRCT1/TORC1 121 Cancer CRK 122 Cancer CRK-II 123 Cancer CRM1 124 Cancer Crry 125 Cancer CSF1R/FMS 126 Cancer CSN5 127 Cancer c-SRC 128 Cancer CATG1B 129 Cancer CTAG2 130 Cancer CTCF 131 Cancer CTFG 132 Cancer CTLA-4 133 Cancer CTNNB1 134 Cancer CTSB 135 Cancer CTSL2 136 Cancer CX3CL1 137 Cancer CXCL12 138 Cancer CYCS 139 Cancer CYLD 140 Cancer CYR61 141 Cancer DAL1L 142 Cancer DAPK1 143 Cancer DBL 144 Cancer DCC 145 Cancer DCN 146 Cancer DCL1 147 Cancer DDB2 148 Cancer DDOST 149 Cancer DDX6 150 Cancer DEK 151 Cancer DHFR 152 Cancer DIABLO 153 Cancer DKK1 154 Cancer DNMT1 155 Cancer DNMT(3A) 156 Cancer DNMT(3B) 157 Cancer DOT1L 158 Cancer DPC4/SMAD4 159 Cancer DPP-IV 160 Cancer E2F 161 Cancer E2F1 162 Cancer E2F1/RBAP 163 Cancer E2F3 164 Cancer EBF1 165 Cancer E-CAD 166 Cancer Ecadherin 167 Cancer EGF 168 Cancer EGFL7 169 Cancer EGFR 170 Cancer EGFR/ERBB-1 171 Cancer EGFR/HER1 172 Cancer EIF4A2 173 Cancer eIF-4E 174 Cancer ELK1 175 Cancer ELK3 176 Cancer EP300 177 Cancer EPCAM 178 Cancer EPH 179 Cancer EPHA1 180 Cancer EPHA3 181 Cancer ER 182 Cancer ERBB-3 183 Cancer ERG 184 Cancer ERK 185 Cancer e-selectin (SELE) 186 Cancer Estrogen Receptor (ESR1) 187 Cancer ETS1 188 Cancer ETS2 189 Cancer ETV6 (TEL) 190 Cancer EZH2 191 Cancer FAK 192 Cancer FANCA 193 Cancer FAP 194 Cancer FAS 195 Cancer FASLG 196 Cancer FBXW7 197 Cancer FER 198 Cancer FGF6 199 Cancer FGF7 200 Cancer FGFR-TACC fusion protein 201 Cancer FGFR1 202 Cancer FGFR2 203 Cancer FGR 204 Cancer Fibroblast growth factor (FGF), 1, 2, 205 Cancer FLI1/ERGB2 206 Cancer FLI1/ERGB2 207 Cancer FLT1 (VEGFR1) 208 Cancer FLT3 209 Cancer FLT4 210 Cancer FMS 211 Cancer FOLH1 (PSMA) 212 Cancer FOS 213 Cancer FOSL1 214 Cancer FOSL2 215 Cancer FOXE1 216 Cancer FPS/FES 217 Cancer FRA1 218 Cancer FRA2 219 Cancer FST 220 Cancer FT3 221 Cancer FUBP1 222 Cancer Furin 223 Cancer FYN 224 Cancer GADD45A 225 Cancer GADD45B 226 Cancer GATA4 227 Cancer GDF2 228 Cancer GIP 229 Cancer GLI 230 Cancer GNA11 231 Cancer GHAQ 232 Cancer GNAS1 233 Cancer GNAS2 234 Cancer GRB-2 235 Cancer GRN 236 Cancer GSK3A 237 Cancer GSP 238 Cancer GST-Pi 239 Cancer HAT1 240 Cancer HCK 241 Cancer HDAC1 242 Cancer HDAC10 243 Cancer HDAC11 244 Cancer HDAC2 245 Cancer HDAC4 246 Cancer HDAC5 247 Cancer HDAC6 248 Cancer HDAC7 249 Cancer HDAC8 250 Cancer HDAC9 251 Cancer Hedgehog 252 Cancer HEK 253 Cancer Her-2 254 Cancer HER2/ERBB2 255 Cancer HER3 256 Cancer HER3/ERBB-2 257 Cancer HER4 258 Cancer HER4/ERBB-4 259 Cancer HIF1A 260 Cancer HIF2A 261 Cancer HIF-1beta 262 Cancer HIND 263 Cancer hMOF 264 Cancer HMGA1 265 Cancer HMGB1 266 Cancer HMTs 267 Cancer HOX11 268 Cancer HOXA7 269 Cancer HOXD10 270 Cancer HPC1 271 Cancer HRAS (c-ha-ras) 272 Cancer HRX/MLLT1 273 Cancer HRX/MLLT2 274 Cancer Hsp27 275 Cancer Hsp70 (HSPBP1) 276 Cancer HSP-90 277 Cancer HST 278 Cancer HST2 279 Cancer HSTF1 280 Cancer HTRA3 281 Cancer IDH (2H) 282 Cancer IDH1 283 Cancer IDH2 284 Cancer IDO 285 Cancer IFNA1 286 Cancer IGF1 287 Cancer IGF1R 288 Cancer IGF2 289 Cancer IGFBP2 290 Cancer IGFBP5 291 Cancer IL-17 292 Cancer IL-23 293 Cancer IL3 294 Cancer IL3RA 295 Cancer IL4RA 296 Cancer IL-6 297 Cancer IL8 298 Cancer ING4 299 Cancer INK4A (p16) 300 Cancer INK4B 301 Cancer INT-1 302 Cancer INT1/WNT1 303 Cancer INT2 304 Cancer IRF1 305 Cancer IRP2 306 Cancer ITGB1 307 Cancer JAG1 308 Cancer JAK1 309 Cancer JAK2 310 Cancer JAK3 311 Cancer JUN 312 Cancer JUNB 313 Cancer JUND 314 Cancer KAT6A 315 Cancer KDM6A 316 Cancer KIF5B 317 Cancer KIP2 318 Cancer KIT 319 Cancer KITLG 320 Cancer KRAS 321 Cancer KRAS2 322 Cancer KRAS2A 323 Cancer KRAS2B 324 Cancer KS3 325 Cancer K-SAM 326 Cancer KSP 327 Cancer LAG3 328 Cancer LATS1 329 Cancer LBC 330 Cancer LCK 331 Cancer LEF1 332 Cancer LET-7 333 Cancer LIMK1 334 Cancer LMO-1 335 Cancer LMO-2 336 Cancer L-MYC 337 Cancer LSD1 338 Cancer 1-selectin 339 Cancer LYL1 340 Cancer LYN 341 Cancer LYT-10 342 Cancer MADH4 343 Cancer MALT1 344 Cancer MAP2K1 345 Cancer MAP3K3 346 Cancer MAP3K10 347 Cancer MAP3K11 348 Cancer MAP3K14 349 Cancer MAP4K4 350 Cancer MAPK 351 Cancer MAPK1 352 Cancer MAPK9 353 Cancer MAS 354 Cancer MAS1 355 Cancer MASXL1 356 Cancer MTA2 357 Cancer MAX 358 Cancer MCC 359 Cancer MCF2 360 Cancer MCL1 361 Cancer MDM2 362 Cancer MDM4 363 Cancer MEF2C 364 Cancer MEK1 365 Cancer MEK2 366 Cancer MEN1 367 Cancer MEN2 368 Cancer MET 369 Cancer Metabolites 370 Cancer Methyltransferase 371 Cancer MGLL 372 Cancer MGMT 373 Cancer MIDHI? 374 Cancer MLH1 375 Cancer MLL 376 Cancer MLLT1/MLL 377 Cancer MLLT2/HRX 378 Cancer MLM 379 Cancer MMP 380 Cancer MMP1 381 Cancer MMP13 382 Cancer MMP2 383 Cancer MMP9 384 Cancer MNK 385 Cancer MOS 386 Cancer MSH2 387 Cancer MSH6 388 Cancer MTG8/RUNX1 389 Cancer MTOR 390 Cancer MTORC2 391 Cancer MUC1 392 Cancer MYB 393 Cancer MYBA 394 Cancer MYBB 395 Cancer MYC (CMYC) 396 Cancer MYCC/MCYN 397 Cancer MYCL1 398 Cancer MYCLK1 399 Cancer MYCN 400 Cancer MYH11//CBFB 401 Cancer MXD1 402 Cancer MXI1 403 Cancer NAFT4 404 Cancer NAFT5 405 Cancer NAIP 406 Cancer Nampt 407 Cancer NANOG 408 Cancer NCL (nucleolin) 409 Cancer NCOA6 410 Cancer NCOR2 411 Cancer NDN 412 Cancer NF1 413 Cancer NF2 414 Cancer NFI-A 415 Cancer NFKB 416 Cancer NFKB1 417 Cancer NFKB2 418 Cancer NGFR 419 Cancer NME1 420 Cancer N-MYC 421 Cancer NOS2A 422 Cancer NOTCH1 423 Cancer NPM1 424 Cancer NPM1/ALK 425 Cancer NPTX1 426 Cancer NR3C1 427 Cancer NRAS 428 Cancer NRG/REL 429 Cancer NSD3 430 Cancer NTRK1 431 Cancer NUAK1 432 Cancer NUP214 433 Cancer OSM 434 Cancer OST 435 Cancer OX40/CD134 436 Cancer P2Y12 437 Cancer P53 (TP53) 438 Cancer P57/KIP2 439 Cancer p85beta 440 Cancer PACE4 441 Cancer PAK4 442 Cancer PALB2 443 Cancer PARP 444 Cancer PARP1 445 Cancer PARP2 446 Cancer PAX-5 447 Cancer PBRM1 448 Cancer PBX1/TCF3 449 Cancer PD1 450 Cancer PDCD4 451 Cancer PDFGR/FILP1L1-PDFGRa 452 Cancer PDGF 453 Cancer PDGFB 454 Cancer PDGFR 455 Cancer PDGFRA 456 Cancer PDL1/2 457 Cancer Pfk 458 Cancer Pfkfb3 459 Cancer PGAM1 460 Cancer PHDGH 461 Cancer PHF6 462 Cancer PI3K 463 Cancer PIGF 464 Cancer PIM1 465 Cancer PKCα 466 Cancer Pkm2 467 Cancer PKN3 468 Cancer PLAU 469 Cancer PLK1 470 Cancer PML/RARA 471 Cancer PMS-1 472 Cancer PMS-2 473 Cancer POLK 474 Cancer POU4F2 475 Cancer PPARD 476 Cancer PPP2CA 477 Cancer PPP2R1A 478 Cancer PPP2R1B 479 Cancer PRAD-1 480 Cancer PRC 481 Cancer PRCA1 482 Cancer Prohibitin 483 Cancer Proteasome inhibitors 484 Cancer PRKCA 485 Cancer PRKRA 486 Cancer PRKG1 487 Cancer PSDK1 488 Cancer P-Selectin 489 Cancer PTCH 490 Cancer PTEN 491 Cancer PTGS2 492 Cancer PTK2B 493 Cancer PTN (pleiotrophin) 494 Cancer RAB6A 495 Cancer RAB6B 496 Cancer RAB21 497 Cancer RAC1 498 Cancer RAC3 499 Cancer RAF 500 Cancer RAF1 501 Cancer RAI 502 Cancer RANKL 503 Cancer RAR-28 504 Cancer RAS 505 Cancer RASL10B (VTS58635) 506 Cancer RRAS 507 Cancer RAASF1 508 Cancer RB1 509 Cancer RBL2 510 Cancer REL 511 Cancer RERG 512 Cancer RET 513 Cancer REST 514 Cancer RFC-1 515 Cancer RHOA 516 Cancer RHOB 517 Cancer RHOBTB2 518 Cancer RHOM-1 519 Cancer RHOM-2 520 Cancer rhPDGF-BB 521 Cancer RNA-R2 522 Cancer ROCK2 523 Cancer ROS1 524 Cancer RTKN 525 Cancer RUNX1 526 Cancer RUNX1/CBFA2T1 527 Cancer RUNXIT1 528 Cancer SDCBP 529 Cancer SEPT9 530 Cancer Ser/Thr 531 Cancer SERPINB5 (MASPIN) 532 Cancer SET 533 Cancer SHC1 534 Cancer SIRT1 535 Cancer SIS 536 Cancer SKI 537 Cancer SLUG 538 Cancer SMAD1 539 Cancer SMAD2 540 Cancer SMAD3 541 Cancer SMAD4 542 Cancer SMAD7 543 Cancer SMARCA4 544 Cancer SFRP1 545 Cancer SKP2 546 Cancer SNAIL 547 Cancer SOCS1 548 Cancer SOS 549 Cancer SOX2 550 Cancer SOX9 551 Cancer SPANXC 552 Cancer SRC1 553 Cancer v-src 554 Cancer SST 555 Cancer STAT1 556 Cancer STAT3 557 Cancer STK11 558 Cancer STX2 559 Cancer Survivin 560 Cancer SYNE1 561 Cancer TACR1 562 Cancer TAL1 563 Cancer TAL2 564 Cancer TAN1 565 Cancer TCF3/PBX1 566 Cancer TCF8/ZEB1 567 Cancer TET2 568 Cancer TFPI2 569 Cancer TFRC (TfR) 570 Cancer TGFB1 571 Cancer TGFB2 572 Cancer TGFBR1 573 Cancer TGFBR2 574 Cancer TGF-α 575 Cancer TGFβ 576 Cancer TGIF2 577 Cancer TGRC 578 Cancer THOC1 579 Cancer THRA1 580 Cancer THRB 581 Cancer TIAM1 582 Cancer TIE2 583 Cancer TIF1A 584 Cancer TIM3/HAVCR2 585 Cancer TIMP1 586 Cancer TIMP2 587 Cancer TIMP3 588 Cancer TIMP4 589 Cancer TK 590 Cancer TLX1 591 Cancer TM1 592 Cancer TMEFF2 593 Cancer TNC 594 Cancer TNFAIP3 595 Cancer TNFα 596 Cancer TNFRSF1A 597 Cancer TNFRSF10A 598 Cancer TNFRSF11A (RANK) 599 Cancer TOP1 600 Cancer TP73L/p63 601 Cancer TPM1 602 Cancer TRIM2 603 Cancer TRK 604 Cancer TRKB 605 Cancer TrkC 606 Cancer TSC1 607 Cancer TSC2 608 Cancer TSG101 609 Cancer Tubulin beta 3 610 Cancer Tubulin beta 5 611 Cancer TUSC2 612 Cancer Twist 613 Cancer TWIST1 614 Cancer Tyr 615 Cancer Tyrosine Kinase Enzymes 616 Cancer VAV 617 Cancer VDR 618 Cancer VCAM 619 Cancer VEGF 620 Cancer VEGFA 621 Cancer VHL 622 Cancer WAF1 623 Cancer WEE1 624 Cancer WIF1 625 Cancer WNT 626 Cancer WNT1 627 Cancer WNT2 628 Cancer WT1 629 Cancer XAF1 630 Cancer XIAP 631 Cancer XPA/XPG 632 Cancer XPO1 633 Cancer YES1 634 Cancer YWHAE 635 Cancer YY1 636 Cancer ZAK (MLT) 637 Cancer ZEB2 638 Cancer αv-β3 639 Cancer RAD51 640 Cancer RAD51C 641 Cancer PPARβ 642 Cancer PPARγ 643 Cancer SPHK2 644 Cancer SPHK1 645 Cancer TMFRSF5B 646 Cancer STAT6 647 Cancer KLF4 648 Cardiovascular/Metabolic Disease ACC 649 Cardiovascular/Metabolic Disease ANGPTL3 650 Cardiovascular/Metabolic Disease Apo(a) 651 Cardiovascular/Metabolic Disease APOA1 652 Cardiovascular/Metabolic Disease APOA4 653 Cardiovascular/Metabolic Disease APOA5 654 Cardiovascular/Metabolic Disease ApoB 655 Cardiovascular/Metabolic Disease ApoB-100 656 Cardiovascular/Metabolic Disease ApoC I 657 Cardiovascular/Metabolic Disease ApoC III 658 Cardiovascular/Metabolic Disease APOE 659 Cardiovascular/Metabolic Disease BACE1 660 Cardiovascular/Metabolic Disease Citrate lyase 661 Cardiovascular/Metabolic Disease DGAT2 662 Cardiovascular/Metabolic Disease endotheal lipase 663 Cardiovascalar/Metabolic Disease Factor VII 664 Cardiovascular/Metabolic Disease Factor IX/F9 665 Cardiovascular/Metabolic Disease FGFR4 666 Cardiovascular/Metabolic Disease GCGR 667 Cardiovascular/Metabolic Disease HDL 668 Cardiovascular/Metabolic Disease LDL 669 Cardiovascular/Metabolic Disease MTTP 670 Cardiovascular/Metabolic Disease PAFAH1B2 671 Cardiovascular/Metabolic Disease PCSK9 672 Cardiovascular/Metabolic Disease PTP-1B 673 Cardiovascalar/Metabolic Disease VLDL Cardiovascular/Metabolic Disease THP—Thrombopoietin for essential thrombocytosis 674 Eye Disease ARMS2 675 Eye Disease CFH 676 Eye Disease C5 677 Eye Disease ERK1 678 Eye Disease ERK2 679 Eye Disease Il-18 680 Eye Disease NGF (proNGF) 681 Eye Disease PDGFC 682 Eye Disease RTP801 683 Eye Disease TLR4 684 Infectious Disease ACEE 685 Infectious Disease aroA 686 Infectious Disease aroC 687 Infectious Disease B2M 688 Infectious Disease carA 689 Infectious Disease CASP1 690 Infectious Disease celB 691 Infectious Disease cflA 692 Infectious Disease cglA 693 Infectious Disease cglE 694 Infectious Disease cilA 695 Infectious Disease cilB 696 Infectious Disease cilC 697 Infectious Disease cilD 698 Infectious Disease cilE 699 Infectious Disease cinA 700 Infectious Disease CCL3 701 Infectious Disease CCL4 702 Infectious Disease CCR5 703 Infectious Disease CD14 704 Infectious Disease CD28 705 Infectious Disease CHIT1 706 Infectious Disease coiA 707 Infectious Disease comA 708 Infectious Disease comC 709 Infectious Disease comX 710 Infectious Disease CSF3 711 Infectious Disease CTL 712 Infectious Disease DDX25 713 Infectious Disease DMC1 714 Infectious Disease Ebola 715 Infectious Disease envZ 716 Infectious Disease epsA 717 Infectious Disease F3 718 Infectious Disease F8 719 Infectious Disease FKBP8 720 Infectious Disease Food borne pathogens 721 Infectious Disease H1N1 722 Infectious Disease H3N2 723 Infectious Disease H5N1 724 Infectious Disease HBx 725 Infectious Disease Hep-A 726 Infectious Disease Hep-B 727 Infectious Disease Hep-C 728 Infectious Disease HIV 729 Infectious Disease HLA-A 730 Infectious Disease HLA-B 731 Infectious Disease HLA-C 732 Infectious Disease HP 733 Infectious Disease HSPD1 734 Infectious Disease IDO1 735 Infectious Disease IL1B 736 Infectious Disease IL6 737 Infectious Disease IL12RB2 738 Infectious Disease IL15 739 Infectious Disease IL17A 740 Infectious Disease IL1RN 741 Infectious Disease Influenza RNA-dependent RNA polymerase 742 Infectious Disease INS 743 Infectious Disease LACTB 744 Infectious Disease LTA 745 Infectious Disease Malaria 746 Infectious Disease MBL2 747 Infectious Disease MIF 748 Infections Disease miR-122 749 Infectious Disease MMP3 750 Infectious Disease NS1A 751 Infectious Disease NS5A 752 Infectious Disease ompF 753 Infectious Disease ostA 754 Infectious Disease pbpG 755 Infectious Disease PPIA 756 Infectious Disease Protease Inhibitors 757 Infectious Disease PRTN3 758 Infectious Disease PTK 759 Infectious Disease PTPRC/CD45 760 Infectious Disease pyrC 761 Infectious Disease relA 762 Infectious Disease retinoic acid receptors/ retinoids 763 Infectious Disease rpmA 764 Infectious Disease rstA 765 Infectious Disease RSV 766 Infectious Disease RSV 767 Infectious Disease SARS 768 Infectious Disease secE 769 Infectious Disease SELL 770 Infectious Disease SERPINA1 771 Infectious Disease SLC11A1 772 Infectious Disease spsC 773 Infectious Disease tcdA 774 Infectious Disease tcdB 775 Infectious Disease TLR2 776 Infectious Disease TLR7 777 Infectious Disease TNF 778 Infectious Disease TNFRSF1B 779 Infectious Disease TNFRSF8 780 Infectious Disease trmD 781 Infectious Disease uppP 782 Infectious Disease West Nile 783 Inflammation ACEI 784 Inflammation ADAMS 785 Inflammation ADAMTS 786 Inflammation AGER 787 Inflammation Aldosterone 788 Inflammation ALK5 789 Inflammation Aminoglycoside 790 Inflammation ARB 791 Inflammation ATG16L1 792 Inflammation BDKRB1 793 Inflammation bFGF 794 Inflammation BMP-7 795 Inflammation c-abl 796 Inflammation CaMKIV 797 Inflammation CASP14 798 Inflammation CCL2/CCL2 receptor 799 Inflammation CCL13 800 Inflammation CCN2 801 Inflammation CCR1 802 Inflammation CCR2 803 Inflammation CCR9 804 Inflammation CCR10 805 Inflammation CD97 806 Inflammation COX 807 Inflammation CRP 808 Inflammation CTGF 809 Inflammation CX3CR1 810 Inflammation CXCR-4 811 Inflammation CXCR-7 812 Inflammation Endothelin 813 Inflammation ELANE 814 Inflammation EPO 815 Inflammation F2RL1 816 Inflammation FPR1 817 Inflammation FPR2 818 Inflammation GPR84 819 Inflammation GZMB 820 Inflammation Hepcidin (HAMP) 821 Inflammation HGF 822 Inflammation HRH4 823 Inflammation ICAM-1 824 Inflammation IFNG 825 Inflammation IL1 826 Inflammation IL10 827 Inflammation IL12 828 Inflammation IL13 829 Inflammation IL2 830 Inflammation IL4 831 Inflammation Il-5 832 Inflammation IL7 833 Inflammation Integrin α4β7 834 Inflammation JNK 835 Inflammation KNG1 836 Inflammation MAPK14 837 Inflammation MCP1 838 Inflammation M-CSF 839 Inflammation MIF1 840 Inflammation MYD88 841 Inflammation Nitric Oxide 842 Inflammation NOD2 843 Inflammation NR1H2 844 Inflammation P38 MAPK 845 Inflammation PAI-1 846 Inflammation PLA2G2D 847 Inflammation PLA2G7 848 Inflammation PLA2G10 849 Inflammation plasminogen 850 Inflammation PLCγ 851 Inflammation PPIG 852 Inflammation PPARα 853 Inflammation PSGL-1 854 Inflammation PTGDR 855 Inflammation PTGDR2 856 Inflammation Rantes (CCL5) 857 Inflammation Renin 858 Inflammation ROCK (Rho-kinase) 859 Inflammation SAA1 860 Inflammation SAP 861 Inflammation SCGB1A1 862 Inflammation SELPLG 863 Inflammation Smads (1, 2, 3, 5) 864 Inflammation SYK 865 Inflammation TLR9 866 Inflammation TSLP 867 Inflammation TNFAIP6 868 Inflammation TNFAIP8L2 869 Inflammation tpa 870 Inflammation uPA 871 Inflammation Vasopeptidase 872 Inflammation VLA-4 873 Inflammation XBP1 874 Neurological Disease alpha-synuclein 875 Neurological Disease ApoE 4 876 Neurological Disease APP 877 Neurological Disease Beta amyloid 878 Neurological Disease CDK5R2 879 Neurological Disease CLU 880 Neurological Disease COX2 881 Neurological Disease CR1 882 Neurological Disease ErbB 883 Neurological Disease FRA10AC1 884 Neurological Disease GBA 885 Neurological Disease GNAS 886 Neurological Disease GPCR 887 Neurological Disease GRM1 888 Neurological Disease GUSB 889 Neurological Disease has-mir-29b 890 Neurological Disease has-mir-29c 891 Neurological Disease HDAC3 892 Neurological Disease hnRNPA1 893 Neurological Disease hnRNPA2B1 894 Neurological Disease hsa-miR-137 895 Neurological Disease HTT 896 Neurological Disease IAPP 897 Neurological Disease LRRK2 898 Neurological Disease MAPT 899 Neurological Disease MBP 900 Neurological Disease MDK (Midkine) 901 Neurological Disease MT-ATP6 902 Neurological Disease PARK 903 Neurological Disease PARK7 904 Neurological Disease PBP 905 Neurological Disease PDE1B 906 Neurological Disease PICALM 907 Neurological Disease PINK1 908 Neurological Disease PON1 909 Neurological Disease PPARGC1B 910 Neurological Disease PRNP 911 Neurological Disease PSEN1 912 Neurological Disease PSEN2 913 Neurological Disease RAGE 914 Neurological Disease SERPINA3 915 Neurological Disease SNCA 916 Neurological Disease SOD1 917 Neurological Disease SPON1 918 Neurological Disease SPP1 919 Neurological Disease STH 920 Neurological Disease Supt4h 921 Neurological Disease Tau 922 Neurological Disease TOMM40 923 Neurological Disease TUBA3 924 Neurological Disease Ubiquilin-2 925 Rare Disease AAT 926 Rare Disease ABCG5 927 Rare Disease ACHE 928 Rare Disease ADA 929 Rare Disease AGXT 930 Rare Disease AIRE 931 Rare Disease ALAS-1 932 Rare Disease ALDH2 933 Rare Disease alpha-1 antritrypsisn 934 Rare Disease AMPH 935 Rare Disease antithrombin 936 Rare Disease AQP2 937 Rare Disease ASPA 938 Rare Disease APT7A 939 Rare Disease ATP7B 940 Rare Disease AVPR2 941 Rare Disease BSCL2 942 Rare Disease C1S 943 Rare Disease CCL3L1 944 Rare Disease CD79A 945 Rare Disease CTLA4 946 Rare Disease CYB5R3 947 Rare Disease CYP117A1 948 Rare Disease CYBB 949 Rare Disease CYP21A2 950 Rare Disease CYP27A1 951 Rare Disease DMPK 952 Rare Disease ENO2 953 Rare Disease F2 954 Rare Disease F5 955 Rare Disease F10 956 Rare Disease FGF23 957 Rare Disease FRAXA 958 Rare Disease FRAXE 959 Rare Disease GAA 960 Rare Disease GAD1 961 Rare Disease GCCR 962 Rare Disease GCK 963 Rare Disease GDNF 964 Rare Disease GFAP 965 Rare Disease GH1 966 Rare Disease GHR 967 Rare Disease GJB1 968 Rare Disease GLA 969 Rare Disease GLRA1 970 Rare Disease GYS2 971 Rare Disease HADHA 972 Rare Disease HFE 973 Rare Disease IGES 974 Rare Disease IPW 975 Rare Disease KCNJ2 976 Rare Disease KRT6A (Keratin K6a) 977 Rare Disease KRT81 978 Rare Disease KRT86 979 Rare Disease LMAN1 980 Rare Disease LMNA 981 Rare Disease MPL 982 Rare Disease MPZ 983 Rare Disease NEU1 984 Rare Disease NPC1 985 Rare Disease NPC2 986 Rare Disease NR0B1 987 Rare Disease NR3C2 988 Rare Disease PKK 989 Rare Disease PMP22 990 Rare Disease PYGM 991 Rare Disease RETN 992 Rare Disease SAG 993 Rare Disease SCNN1A 994 Rare Disease SH2D1A 995 Rare Disease SLC2A1 (Glut1) 996 Rare Disease SMN2 997 Rare Disease SMPD1 998 Rare Disease SNRPN 999 Rare Disease THBD 1000 Rare Disease STAR 1001 Rare Disease SYP 1002 Rare Disease TRD 1003 Rare Disease TSHB 1004 Rare Disease Tmprss6 1005 Rare Disease TTR 1006 Rare Disease UBE3A 1007 Rare Disease WAS 1008 Rare Disease WRN 1009 Rare Disease Dentatorubropallidoluysian Atrophy 1010 Rare Disease Huntington′s Disease 1011 Rare Disease Spinobulbar Muscular Atrophy 1012 Rare Disease SCA1 (Spinocerebellar Ataxia Type 1) 1013 Rare Disease SCA2 (Spinocerebellar Ataxia Type 2) 1014 Rare Disease SCA3 (Spinocerebellar Ataxia Type 3 or Machado-Joseph Disease) 1015 Rare Disease SCA6 (Spinocerebellar Ataxia Type 6) 1016 Rare Disease SCA7 (Spinocerebellar Ataxia Type 7) 1017 Rare Disease Fragile X Syndrome 1018 Rare Disease Fragile XE Mental Retardation 1019 Rare Disease Friedreich′s Ataxia 1020 Rare Disease Myotonic Dystrophy 1021 Rare Disease Spinocerebellar Ataxia Type 8 1022 Rare Disease Spinocerebellar Ataxia Type 12 1023 Rare Disease SPT4 1024 Rare Disease ATN1 1025 Rare Disease DRPLA 1026 Rare Disease HTT 1027 Rare Disease ATXN1 1028 Rare Disease ATXN2 1029 Rare Disease ATXN3 1030 Rare Disease CACNA1A 1031 Rare Disease ATXN7 1032 Rare Disease TBP 1033 Rare Disease FMR1 1034 Rare Disease AFF2 1035 Rare Disease FXN 1036 Rare Disease SCA8 1037 Rare Disease PPP2R2B 1038 Stem Cells Cancer Stem Cells 1039 Stem Cells Cardiac Stem Cells 1040 Stem Cells Kidney Stem Cells 1041 Stem Cells Embryonic Stem Cells 1042 Stem Cells Tissue Stem Cells 1043 Stem Cells Induced Pluripotent Stem Cells 1044 Stem Cells Blood Stem Cells 1045 Stem Cells Mescenchymal Stem Cells 1046 Stem Cells Cord Blood Stem Cells

Non-Cancer Targets

The present invention is not limited to the targeting of cancer genes. The methods and compositions of the present invention find use in the targeting of any gene that it is desirable to down regulate the expression of. For example, targets for immune and/or surface antigens or immune surveillance targets, angiogenic receptors, proteins and factors (kinases, heat shock, hypoxic, oxidative stress gene/protein targets), monogenic diseases, inflammation, gene transcription (transcription factors, cis regulatory elements), cell recognition receptors, cell signaling receptors, cell death (autophagy, necrosis, apoptosis), cell adhesion, survival targets (resistance), metastases targets (brain, primary to secondary tumors), chemokines/cytokines, EMT/MET, immune cell activation factors, multidrug resistance, viral proteins and viral recognition proteins, psoriasis, dermatitis and eczema

Extracellular matrix, stromal or connective tissue genes/proteins, coagulation factors and platelet aggregation or platelet overproduction, and growth factors.

For example, in some embodiments, the genes to be targeted include, but are not limited to, an immunoglobulin or antibody gene, a clotting factor gene, a protease, a pituitary hormone, a protease inhibitor, a growth factor, a somatomedian, a gonadotrophin, a chemotactin, a chemokine, a plasma protein, a plasma protease inhibitor, an interleukin, an interferon, a cytokine, a transcription factor, or a pathogen target (e.g., a viral gene, a bacterial gene, a microbial gene, a fungal gene).

In other embodiments and gene from a pathogen is targeted. Exemplary pathogens include, but are not limited to, Human Immunodeficiency virus (CD4, APOBEC3G, Vif, LEDGF/p75), Hepatitis B virus, hepatitis C virus (SR-B1, scavenger receptor type B1; CLDN-1, claudin-1; OCLN, occluding), hepatitis A virus, respiratory syncytial virus, pathogens involved in severe acute respiratory syndrome, west nile virus, and food borne pathogens (e.g., E. coli).

The lists of Cancer and Non-Cancer targets from above is intended to be specific and accurate, but in addition to the targets above we have further found and we describe in even greater detail the targets listed below, comprising both cancer and non-cancer targets, presented in no particular order. These targets are especially well suited for DNAi targeting and therapy. The preferred list of targets is provided with the sections that follow which provided detailed descriptions of over 40 genes. These gene targets are numbered below, 1-30. Included with a description of many of these preferred targets are the background relevance of the gene, gene identification, the targeted oligonucleotide sequences, the hot zones, and the 5′ upstream genetic code.

EXPERIMENTALS

These examples are provided in order to demonstrate and further illustrate certain preferred embodiments and aspects of the present invention and are not to be construed as limiting the scope thereof.

In the experimental disclosure which follows, the following abbreviations apply: N (normal); M (molar); mM (millimolar); μM (micromolar); mol (moles); mmol (millimoles); μmol (micromoles); nmol (nanomoles); pmol (picomoles); g (grams); mg (milligrams); μg (micrograms); ng (nanograms); 1 or L (liters); ml (milliliters); μl (microliters); cm (centimeters); mm (millimeters); μm (micrometers); nm (nanometers); and ° C. (degrees Centigrade).

1) Survivin. Survivin (BIRC5) also called buloviral inhibitor of apoptosis repeat-containing 5 is a member of the inhibitor of apoptosis family that is expressed during mitosis in a cell cycle-dependent manner. Survivin is localized to different components of the mitotic apparatus, plays an important role in both cell division and inhibition of apoptosis. Survivin is not expressed in normal adult tissue, but is widely expressed in a majority of cancers (Fukuda and Pelus, Mol Cancer Ther 2006; 5 1087-1098), often with poor prognosis. Survivin inhibits caspase activation, the key effector enzyme in programmed cell death, and as a result there is uncontrolled growth and drug resistance. The inhibition of survivin leads to increased apoptosis and decreased tumor growth and sensitizes cells to various therapeutic interventions including chemotherapies and targeted therapies against cancer targets. Survivin expression is increased in tumors and regulated by the cell cycle (expressed in mitosis in a cell cycle dependent manner); expression is also linked to p53 and is targeted by the WNT1 pathway and is upregulated by β-catenin. A review of approaches targeted against survivin may be found in “Targeting surviving in cancer: a patent review” (Expert Opinion on Therapeutic Patents, December 2010, Vol. 20, No. 12: Pages 1723-1737).

An antisense therapeutic being developed (LY2181308) downregulates survivin expression in human cancer cells derived from lung, colon, pancreas, liver, breast, prostate, ovary, cervix, skin, and brain as measured by quantitative RT-PCR and immunoblotting analysis (Carrasco et al., Mol Cancer Ther 2011; 10(2); 221-32). Specific inhibition of survivin expression in multiple cancer cell lines induced caspase-3-dependent apoptosis, cell cycle arrest in the G2-M phase, and multinucleated cells and sensitized tumor cells to chemotherapeutic-induced apoptosis. In an in vivo human xenograft tumor model, LY2181308 produced significant antitumor activity as compared with saline or its sequence-specific control oligonucleotide and sensitized to gemcitabine, paclitaxel, and docetaxel with inhibition of surviving expression in xenograft tumors. LY2181308 is being evaluated in a clinical setting (Phase II) in combination with docetaxel for the treatment of prostate cancer.

Protein: Survivin Gene: BIRC5 (Homo sapiens, chromosome 17, 76210277-76221716 [NCBI Reference Sequence: NC_(—)000017.10]; start site location: 76210398; strand: positive)

Gene Identification GeneID 332 HGNC 593 HPRD 04520 MIM 603352

Targeted Sequences Relative upstream location Se- to gene quence Design start ID No: ID Sequence (5′-3′) site 1 SU1 GAGCGCACGCCCTCTTAGGCGG 73 75 SU2 CACCCCGAGGTACGATCAGTGCGTACC 2990 105 SU3 GACATCGCTGTCCCGGCGAGTACATCGTT 665 155 SU1_02 GAGCGCACGCCCTCTTAGGCG 73 229 SU1_03 GAGCGCACGCCCTCTTAGGCGGTCCA 73 303 GTCGCCCCTGGGTCCTGCTGATTGGC 1918 322 CAGCGAGCCTGGGCCCCATCGGCACATCT 2905 357 CCCGCGGCCTTCTGGGAGTAGAGGC 102 431 TCCCGGCGAGTACATCGTTGACTGCACG 675 481 AACCTCCTCCCCGCCACGGGTT 1229

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 1 GAGCGCACGCCCTCTTAGGCGG 77 2 AGCGCACGCCCTCTTAGGCG 78 3 GCGCACGCCCTCTTAGGCGG 79 4 CGCACGCCCTCTTAGGCGGT 80 5 GCACGCCCTCTTAGGCGGTC 81 6 CACGCCCTCTTAGGCGGTCC 82 7 ACGCCCTCTTAGGCGGTCCA 83 8 CGCCCTCTTAGGCGGTCCAC 84 9 GCCCTCTTAGGCGGTCCACC 85 10 CCCTCTTAGGCGGTCCACCC 86 11 CCTCTTAGGCGGTCCACCCC 87 12 CTCTTAGGCGGTCCACCCCC 88 13 TCTTAGGCGGTCCACCCCCC 89 14 CTTAGGCGGTCCACCCCCCG 90 15 TTAGGCGGTCCACCCCCCGC 91 16 TAGGCGGTCCACCCCCCGCG 92 17 AGGCGGTCCACCCCCCGCGG 93 18 GGCGGTCCACCCCCCGCGGC 94 19 GCGGTCCACCCCCCGCGGCC 95 20 CGGTCCACCCCCCGCGGCCT 96 21 GGTCCACCCCCCGCGGCCTT 97 22 GTCCACCCCCCGCGGCCTTC 98 23 TCCACCCCCCGCGGCCTTCT 99 24 CCACCCCCCGCGGCCTTCTG 100 25 CACCCCCCGCGGCCTTCTGG 101 26 ACCCCCCGCGGCCTTCTGGG 102 27 CCCCCCGCGGCCTTCTGGGA 103 28 CCCCCGCGGCCTTCTGGGAG 104 29 CCCCGCGGCCTTCTGGGAGT 105 30 CCCGCGGCCTTCTGGGAGTA 106 31 CCGCGGCCTTCTGGGAGTAG 107 32 CGCGGCCTTCTGGGAGTAGA 108 33 GCGGCCTTCTGGGAGTAGAG 109 34 CGGCCTTCTGGGAGTAGAGG 110 35 GGAGCGCACGCCCTCTTAGG 76 36 GGGAGCGCACGCCCTCTTAG 75 37 CGGGAGCGCACGCCCTCTTA 74 38 TCGGGAGCGCACGCCCTCTT 73 39 GTCGGGAGCGCACGCCCTCT 72 40 TGTCGGGAGCGCACGCCCTC 71 41 ATGTCGGGAGCGCACGCCCT 70 42 CATGTCGGGAGCGCACGCCC 69 43 GCATGTCGGGAGCGCACGCC 68 44 GGCATGTCGGGAGCGCACGC 67 45 GGGCATGTCGGGAGCGCACG 66 46 GGGGCATGTCGGGAGCGCAC 65 47 CGGGGCATGTCGGGAGCGCA 64 48 GCGGGGCATGTCGGGAGCGC 63 49 CGCGGGGCATGTCGGGAGCG 62 50 CCGCGGGGCATGTCGGGAGC 61 51 GCCGCGGGGCATGTCGGGAG 60 52 CGCCGCGGGGCATGTCGGGA 59 53 GCGCCGCGGGGCATGTCGGG 58 54 CGCGCCGCGGGGCATGTCGG 57 55 GCGCGCCGCGGGGCATGTCG 56 56 GGCGCGCCGCGGGGCATGTC 55 57 TGGCGCGCCGCGGGGCATGT 54 58 ATGGCGCGCCGCGGGGCATG 53 59 AATGGCGCGCCGCGGGGCAT 52 60 TAATGGCGCGCCGCGGGGCA 51 61 TTAATGGCGCGCCGCGGGGC 50 62 GTTAATGGCGCGCCGCGGGG 49 63 GGTTAATGGCGCGCCGCGGG 48 64 CGGTTAATGGCGCGCCGCGG 47 65 GCGGTTAATGGCGCGCCGCG 46 66 GGCGGTTAATGGCGCGCCGC 45 67 TGGCGGTTAATGGCGCGCCG 44 68 CTGGCGGTTAATGGCGCGCC 43 69 TCTGGCGGTTAATGGCGCGC 42 70 ATCTGGCGGTTAATGGCGCG 41 71 AATCTGGCGGTTAATGGCGC 40 72 AAATCTGGCGGTTAATGGCG 39 73 CAAATCTGGCGGTTAATGGC 38 74 TCAAATCTGGCGGTTAATGG 37 75 CACCCCGAGGTACGATCAGTGCGTACC 2994 76 ACCCCGAGGTACGATCAGTG 2995 77 CCCCGAGGTACGATCAGTGC 2996 78 CCCGAGGTACGATCAGTGCG 2997 79 CCGAGGTACGATCAGTGCGT 2998 80 CGAGGTACGATCAGTGCGTA 2999 81 GAGGTACGATCAGTGCGTAC 3000 82 AGGTACGATCAGTGCGTACC 3001 83 GGTACGATCAGTGCGTACCA 3002 84 GTACGATCAGTGCGTACCAA 3003 85 TACGATCAGTGCGTACCAAG 3004 86 ACGATCAGTGCGTACCAAGT 3005 87 CGATCAGTGCGTACCAAGTA 3006 88 GATCAGTGCGTACCAAGTAC 3007 89 ATCAGTGCGTACCAAGTACA 3008 90 TCAGTGCGTACCAAGTACAT 3009 91 CAGTGCGTACCAAGTACATA 3010 92 CCACCCCGAGGTACGATCAG 2993 93 CCCACCCCGAGGTACGATCA 2992 94 TCCCACCCCGAGGTACGATC 2991 95 CTCCCACCCCGAGGTACGAT 2990 96 TCTCCCACCCCGAGGTACGA 2989 97 TTCTCCCACCCCGAGGTACG 2988 98 CTTCTCCCACCCCGAGGTAC 2987 99 TCTTCTCCCACCCCGAGGTA 2986 100 CTCTTCTCCCACCCCGAGGT 2985 101 TCTCTTCTCCCACCCCGAGG 2984 102 CTCTCTTCTCCCACCCCGAG 2983 103 CCTCTCTTCTCCCACCCCGA 2982 104 CCCTCTCTTCTCCCACCCCG 2981 105 GACATCGCTGTCCCGGCGAGTACATCGTT 669 106 ACATCGCTGTCCCGGCGAGT 670 107 CATCGCTGTCCCGGCGAGTA 671 108 ATCGCTGTCCCGGCGAGTAC 672 109 TCGCTGTCCCGGCGAGTACA 673 110 CGCTGTCCCGGCGAGTACAT 674 111 GCTGTCCCGGCGAGTACATC 675 112 CTGTCCCGGCGAGTACATCG 676 113 TGTCCCGGCGAGTACATCGT 677 114 GTCCCGGCGAGTACATCGTT 678 115 TCCCGGCGAGTACATCGTTG 679 116 CCCGGCGAGTACATCGTTGA 680 117 CCGGCGAGTACATCGTTGAC 681 118 CGGCGAGTACATCGTTGACT 682 119 GGCGAGTACATCGTTGACTG 683 120 GCGAGTACATCGTTGACTGC 684 121 CGAGTACATCGTTGACTGCA 685 122 GAGTACATCGTTGACTGCAC 686 123 AGTACATCGTTGACTGCACG 687 124 GTACATCGTTGACTGCACGA 688 125 TACATCGTTGACTGCACGAC 689 126 ACATCGTTGACTGCACGACC 690 127 CATCGTTGACTGCACGACCT 691 128 ATCGTTGACTGCACGACCTG 692 129 TCGTTGACTGCACGACCTGG 693 130 CGTTGACTGCACGACCTGGG 694 131 GTTGACTGCACGACCTGGGT 695 132 TTGACTGCACGACCTGGGTT 696 133 TGACTGCACGACCTGGGTTT 697 134 GACTGCACGACCTGGGTTTC 698 135 ACTGCACGACCTGGGTTTCC 699 136 CTGCACGACCTGGGTTTCCA 700 137 TGCACGACCTGGGTTTCCAG 701 138 GCACGACCTGGGTTTCCAGG 702 139 CACGACCTGGGTTTCCAGGA 703 140 ACGACCTGGGTTTCCAGGAG 704 141 CGACCTGGGTTTCCAGGAGG 705 142 AGACATCGCTGTCCCGGCGA 668 143 CAGACATCGCTGTCCCGGCG 667 144 GCAGACATCGCTGTCCCGGC 666 145 AGCAGACATCGCTGTCCCGG 665 146 CAGCAGACATCGCTGTCCCG 664 147 GCAGCAGACATCGCTGTCCC 663 148 TGCAGCAGACATCGCTGTCC 662 149 GTGCAGCAGACATCGCTGTC 661 150 AGTGCAGCAGACATCGCTGT 660 151 GAGTGCAGCAGACATCGCTG 659 152 GGAGTGCAGCAGACATCGCT 658 153 TGGAGTGCAGCAGACATCGC 657 154 ATGGAGTGCAGCAGACATCG 656 155 GAGCGCACGCCCTCTTAGGCG 77 156 AGCGCACGCCCTCTTAGGCG 78 157 GCGCACGCCCTCTTAGGCGG 79 158 CGCACGCCCTCTTAGGCGGT 80 159 GCACGCCCTCTTAGGCGGTC 81 160 CACGCCCTCTTAGGCGGTCC 82 161 ACGCCCTCTTAGGCGGTCCA 83 162 CGCCCTCTTAGGCGGTCCAC 84 163 GCCCTCTTAGGCGGTCCACC 85 164 CCCTCTTAGGCGGTCCACCC 86 165 CCTCTTAGGCGGTCCACCCC 87 166 CTCTTAGGCGGTCCACCCCC 88 167 TCTTAGGCGGTCCACCCCCC 89 168 CTTAGGCGGTCCACCCCCCG 90 169 TTAGGCGGTCCACCCCCCGC 91 170 TAGGCGGTCCACCCCCCGCG 92 171 AGGCGGTCCACCCCCCGCGG 93 172 GGCGGTCCACCCCCCGCGGC 94 173 GCGGTCCACCCCCCGCGGCC 95 174 CGGTCCACCCCCCGCGGCCT 96 175 GGTCCACCCCCCGCGGCCTT 97 176 GTCCACCCCCCGCGGCCTTC 98 177 TCCACCCCCCGCGGCCTTCT 99 178 CCACCCCCCGCGGCCTTCTG 100 179 CACCCCCCGCGGCCTTCTGG 101 180 ACCCCCCGCGGCCTTCTGGG 102 181 CCCCCCGCGGCCTTCTGGGA 103 182 CCCCCGCGGCCTTCTGGGAG 104 183 CCCCGCGGCCTTCTGGGAGT 105 184 CCCGCGGCCTTCTGGGAGTA 106 185 CCGCGGCCTTCTGGGAGTAG 107 186 CGCGGCCTTCTGGGAGTAGA 108 187 GCGGCCTTCTGGGAGTAGAG 109 188 CGGCCTTCTGGGAGTAGAGG 110 189 GGAGCGCACGCCCTCTTAGG 76 190 GGGAGCGCACGCCCTCTTAG 75 191 CGGGAGCGCACGCCCTCTTA 74 192 TCGGGAGCGCACGCCCTCTT 73 193 GTCGGGAGCGCACGCCCTCT 72 194 TGTCGGGAGCGCACGCCCTC 71 195 ATGTCGGGAGCGCACGCCCT 70 196 CATGTCGGGAGCGCACGCCC 69 197 GCATGTCGGGAGCGCACGCC 68 198 GGCATGTCGGGAGCGCACGC 67 199 GGGCATGTCGGGAGCGCACG 66 200 GGGGCATGTCGGGAGCGCAC 65 201 CGGGGCATGTCGGGAGCGCA 64 202 GCGGGGCATGTCGGGAGCGC 63 203 CGCGGGGCATGTCGGGAGCG 62 204 CCGCGGGGCATGTCGGGAGC 61 205 GCCGCGGGGCATGTCGGGAG 60 206 CGCCGCGGGGCATGTCGGGA 59 207 GCGCCGCGGGGCATGTCGGG 58 208 CGCGCCGCGGGGCATGTCGG 57 209 GCGCGCCGCGGGGCATGTCG 56 210 GGCGCGCCGCGGGGCATGTC 55 211 TGGCGCGCCGCGGGGCATGT 54 212 ATGGCGCGCCGCGGGGCATG 53 213 AATGGCGCGCCGCGGGGCAT 52 214 TAATGGCGCGCCGCGGGGCA 51 215 TTAATGGCGCGCCGCGGGGC 50 216 GTTAATGGCGCGCCGCGGGG 49 217 GGTTAATGGCGCGCCGCGGG 48 218 CGGTTAATGGCGCGCCGCGG 47 219 GCGGTTAATGGCGCGCCGCG 46 220 GGCGGTTAATGGCGCGCCGC 45 221 TGGCGGTTAATGGCGCGCCG 44 222 CTGGCGGTTAATGGCGCGCC 43 223 TCTGGCGGTTAATGGCGCGC 42 224 ATCTGGCGGTTAATGGCGCG 41 225 AATCTGGCGGTTAATGGCGC 40 226 AAATCTGGCGGTTAATGGCG 39 227 CAAATCTGGCGGTTAATGGC 38 228 TCAAATCTGGCGGTTAATGG 37 229 GAGCGCACGCCCTCTTAGGCGGTCCA 77 230 AGCGCACGCCCTCTTAGGCG 78 231 GCGCACGCCCTCTTAGGCGG 79 232 CGCACGCCCTCTTAGGCGGT 80 233 GCACGCCCTCTTAGGCGGTC 81 234 CACGCCCTCTTAGGCGGTCC 82 235 ACGCCCTCTTAGGCGGTCCA 83 236 CGCCCTCTTAGGCGGTCCAC 84 237 GCCCTCTTAGGCGGTCCACC 85 238 CCCTCTTAGGCGGTCCACCC 86 239 CCTCTTAGGCGGTCCACCCC 87 240 CTCTTAGGCGGTCCACCCCC 88 241 TCTTAGGCGGTCCACCCCCC 89 242 CTTAGGCGGTCCACCCCCCG 90 243 TTAGGCGGTCCACCCCCCGC 91 244 TAGGCGGTCCACCCCCCGCG 92 245 AGGCGGTCCACCCCCCGCGG 93 246 GGCGGTCCACCCCCCGCGGC 94 247 GCGGTCCACCCCCCGCGGCC 95 248 CGGTCCACCCCCCGCGGCCT 96 249 GGTCCACCCCCCGCGGCCTT 97 250 GTCCACCCCCCGCGGCCTTC 98 251 TCCACCCCCCGCGGCCTTCT 99 252 CCACCCCCCGCGGCCTTCTG 100 253 CACCCCCCGCGGCCTTCTGG 101 254 ACCCCCCGCGGCCTTCTGGG 102 255 CCCCCCGCGGCCTTCTGGGA 103 256 CCCCCGCGGCCTTCTGGGAG 104 257 CCCCGCGGCCTTCTGGGAGT 105 258 CCCGCGGCCTTCTGGGAGTA 106 259 CCGCGGCCTTCTGGGAGTAG 107 260 CGCGGCCTTCTGGGAGTAGA 108 261 GCGGCCTTCTGGGAGTAGAG 109 262 CGGCCTTCTGGGAGTAGAGG 110 263 GGAGCGCACGCCCTCTTAGG 76 264 GGGAGCGCACGCCCTCTTAG 75 265 CGGGAGCGCACGCCCTCTTA 74 266 TCGGGAGCGCACGCCCTCTT 73 267 GTCGGGAGCGCACGCCCTCT 72 268 TGTCGGGAGCGCACGCCCTC 71 269 ATGTCGGGAGCGCACGCCCT 70 270 CATGTCGGGAGCGCACGCCC 69 271 GCATGTCGGGAGCGCACGCC 68 272 GGCATGTCGGGAGCGCACGC 67 273 GGGCATGTCGGGAGCGCACG 66 274 GGGGCATGTCGGGAGCGCAC 65 275 CGGGGCATGTCGGGAGCGCA 64 276 GCGGGGCATGTCGGGAGCGC 63 277 CGCGGGGCATGTCGGGAGCG 62 278 CCGCGGGGCATGTCGGGAGC 61 279 GCCGCGGGGCATGTCGGGAG 60 280 CGCCGCGGGGCATGTCGGGA 59 281 GCGCCGCGGGGCATGTCGGG 58 282 CGCGCCGCGGGGCATGTCGG 57 283 GCGCGCCGCGGGGCATGTCG 56 284 GGCGCGCCGCGGGGCATGTC 55 285 TGGCGCGCCGCGGGGCATGT 54 286 ATGGCGCGCCGCGGGGCATG 53 287 AATGGCGCGCCGCGGGGCAT 52 288 TAATGGCGCGCCGCGGGGCA 51 289 TTAATGGCGCGCCGCGGGGC 50 290 GTTAATGGCGCGCCGCGGGG 49 291 GGTTAATGGCGCGCCGCGGG 48 292 CGGTTAATGGCGCGCCGCGG 47 293 GCGGTTAATGGCGCGCCGCG 46 294 GGCGGTTAATGGCGCGCCGC 45 295 TGGCGGTTAATGGCGCGCCG 44 296 CTGGCGGTTAATGGCGCGCC 43 297 TCTGGCGGTTAATGGCGCGC 42 298 ATCTGGCGGTTAATGGCGCG 41 299 AATCTGGCGGTTAATGGCGC 40 300 AAATCTGGCGGTTAATGGCG 39 301 CAAATCTGGCGGTTAATGGC 38 302 TCAAATCTGGCGGTTAATGG 37 303 GTCGCCCCTGGGTCCTGCTGATTGGC 1919 304 TCGCCCCTGGGTCCTGCTGA 1920 305 CGCCCCTGGGTCCTGCTGAT 1921 306 GGTCGCCCCTGGGTCCTGCT 1918 307 AGGTCGCCCCTGGGTCCTGC 1917 308 CAGGTCGCCCCTGGGTCCTG 1916 309 GCAGGTCGCCCCTGGGTCCT 1915 310 GGCAGGTCGCCCCTGGGTCC 1914 311 TGGCAGGTCGCCCCTGGGTC 1913 312 TTGGCAGGTCGCCCCTGGGT 1912 313 TTTGGCAGGTCGCCCCTGGG 1911 314 CTTTGGCAGGTCGCCCCTGG 1910 315 ACTTTGGCAGGTCGCCCCTG 1909 316 GACTTTGGCAGGTCGCCCCT 1908 317 TGACTTTGGCAGGTCGCCCC 1907 318 TTGACTTTGGCAGGTCGCCC 1906 319 GTTGACTTTGGCAGGTCGCC 1905 320 AGTTGACTTTGGCAGGTCGC 1904 321 CAGTTGACTTTGGCAGGTCG 1903 322 CAGCGAGCCTGGGCCCCATCGGCACATCT 2909 323 AGCGAGCCTGGGCCCCATCG 2910 324 GCGAGCCTGGGCCCCATCGG 2911 325 CGAGCCTGGGCCCCATCGGC 2912 326 GAGCCTGGGCCCCATCGGCA 2913 327 AGCCTGGGCCCCATCGGCAC 2914 328 GCCTGGGCCCCATCGGCACA 2915 329 CCTGGGCCCCATCGGCACAT 2916 330 CTGGGCCCCATCGGCACATC 2917 331 TGGGCCCCATCGGCACATCT 2918 332 GGGCCCCATCGGCACATCTG 2919 333 GGCCCCATCGGCACATCTGA 2920 334 GCCCCATCGGCACATCTGAA 2921 335 CCCCATCGGCACATCTGAAG 2922 336 CCCATCGGCACATCTGAAGG 2923 337 CCATCGGCACATCTGAAGGT 2924 338 CATCGGCACATCTGAAGGTG 2925 339 ATCGGCACATCTGAAGGTGC 2926 340 TCGGCACATCTGAAGGTGCA 2927 341 CGGCACATCTGAAGGTGCAC 2928 342 GCAGCGAGCCTGGGCCCCAT 2908 343 TGCAGCGAGCCTGGGCCCCA 2907 344 CTGCAGCGAGCCTGGGCCCC 2906 345 TCTGCAGCGAGCCTGGGCCC 2905 346 ATCTGCAGCGAGCCTGGGCC 2904 347 CATCTGCAGCGAGCCTGGGC 2903 348 CCATCTGCAGCGAGCCTGGG 2902 349 GCCATCTGCAGCGAGCCTGG 2901 350 GGCCATCTGCAGCGAGCCTG 2900 351 GGGCCATCTGCAGCGAGCCT 2899 352 GGGGCCATCTGCAGCGAGCC 2898 353 GGGGGCCATCTGCAGCGAGC 2897 354 AGGGGGCCATCTGCAGCGAG 2896 355 AAGGGGGCCATCTGCAGCGA 2895 356 GAAGGGGGCCATCTGCAGCG 2894 357 CCCGCGGCCTTCTGGGAGTAGAGGC 106 358 CCGCGGCCTTCTGGGAGTAG 107 359 CGCGGCCTTCTGGGAGTAGA 108 360 GCGGCCTTCTGGGAGTAGAG 109 361 CGGCCTTCTGGGAGTAGAGG 110 362 CCCCGCGGCCTTCTGGGAGT 105 363 CCCCCGCGGCCTTCTGGGAG 104 364 CCCCCCGCGGCCTTCTGGGA 103 365 ACCCCCCGCGGCCTTCTGGG 102 366 CACCCCCCGCGGCCTTCTGG 101 367 CCACCCCCCGCGGCCTTCTG 100 368 TCCACCCCCCGCGGCCTTCT 99 369 GTCCACCCCCCGCGGCCTTC 98 370 GGTCCACCCCCCGCGGCCTT 97 371 CGGTCCACCCCCCGCGGCCT 96 372 GCGGTCCACCCCCCGCGGCC 95 373 GGCGGTCCACCCCCCGCGGC 94 374 AGGCGGTCCACCCCCCGCGG 93 375 TAGGCGGTCCACCCCCCGCG 92 376 TTAGGCGGTCCACCCCCCGC 91 377 CTTAGGCGGTCCACCCCCCG 90 378 TCTTAGGCGGTCCACCCCCC 89 379 CTCTTAGGCGGTCCACCCCC 88 380 CCTCTTAGGCGGTCCACCCC 87 381 CCCTCTTAGGCGGTCCACCC 86 382 GCCCTCTTAGGCGGTCCACC 85 383 CGCCCTCTTAGGCGGTCCAC 84 384 ACGCCCTCTTAGGCGGTCCA 83 385 CACGCCCTCTTAGGCGGTCC 82 386 GCACGCCCTCTTAGGCGGTC 81 387 CGCACGCCCTCTTAGGCGGT 80 388 GCGCACGCCCTCTTAGGCGG 79 389 AGCGCACGCCCTCTTAGGCG 78 390 GAGCGCACGCCCTCTTAGGC 77 391 GGAGCGCACGCCCTCTTAGG 76 392 GGGAGCGCACGCCCTCTTAG 75 393 CGGGAGCGCACGCCCTCTTA 74 394 TCGGGAGCGCACGCCCTCTT 73 395 GTCGGGAGCGCACGCCCTCT 72 396 TGTCGGGAGCGCACGCCCTC 71 397 ATGTCGGGAGCGCACGCCCT 70 398 CATGTCGGGAGCGCACGCCC 69 399 GCATGTCGGGAGCGCACGCC 68 400 GGCATGTCGGGAGCGCACGC 67 401 GGGCATGTCGGGAGCGCACG 66 402 GGGGCATGTCGGGAGCGCAC 65 403 CGGGGCATGTCGGGAGCGCA 64 404 GCGGGGCATGTCGGGAGCGC 63 405 CGCGGGGCATGTCGGGAGCG 62 406 CCGCGGGGCATGTCGGGAGC 61 407 GCCGCGGGGCATGTCGGGAG 60 408 CGCCGCGGGGCATGTCGGGA 59 409 GCGCCGCGGGGCATGTCGGG 58 410 CGCGCCGCGGGGCATGTCGG 57 411 GCGCGCCGCGGGGCATGTCG 56 412 GGCGCGCCGCGGGGCATGTC 55 413 TGGCGCGCCGCGGGGCATGT 54 414 ATGGCGCGCCGCGGGGCATG 53 415 AATGGCGCGCCGCGGGGCAT 52 416 TAATGGCGCGCCGCGGGGCA 51 417 TTAATGGCGCGCCGCGGGGC 50 418 GTTAATGGCGCGCCGCGGGG 49 419 GGTTAATGGCGCGCCGCGGG 48 420 CGGTTAATGGCGCGCCGCGG 47 421 GCGGTTAATGGCGCGCCGCG 46 422 GGCGGTTAATGGCGCGCCGC 45 423 TGGCGGTTAATGGCGCGCCG 44 424 CTGGCGGTTAATGGCGCGCC 43 425 TCTGGCGGTTAATGGCGCGC 42 426 ATCTGGCGGTTAATGGCGCG 41 427 AATCTGGCGGTTAATGGCGC 40 428 AAATCTGGCGGTTAATGGCG 39 429 CAAATCTGGCGGTTAATGGC 38 430 TCAAATCTGGCGGTTAATGG 37 431 TCCCGGCGAGTACATCGTTGACTGCACG 679 432 CCCGGCGAGTACATCGTTGA 680 433 CCGGCGAGTACATCGTTGAC 681 434 CGGCGAGTACATCGTTGACT 682 435 GGCGAGTACATCGTTGACTG 683 436 GCGAGTACATCGTTGACTGC 684 437 CGAGTACATCGTTGACTGCA 685 438 GAGTACATCGTTGACTGCAC 686 439 AGTACATCGTTGACTGCACG 687 440 GTACATCGTTGACTGCACGA 688 441 TACATCGTTGACTGCACGAC 689 442 ACATCGTTGACTGCACGACC 690 443 CATCGTTGACTGCACGACCT 691 444 ATCGTTGACTGCACGACCTG 692 445 TCGTTGACTGCACGACCTGG 693 446 CGTTGACTGCACGACCTGGG 694 447 GTTGACTGCACGACCTGGGT 695 448 TTGACTGCACGACCTGGGTT 696 449 TGACTGCACGACCTGGGTTT 697 450 GACTGCACGACCTGGGTTTC 698 451 ACTGCACGACCTGGGTTTCC 699 452 CTGCACGACCTGGGTTTCCA 700 453 TGCACGACCTGGGTTTCCAG 701 454 GCACGACCTGGGTTTCCAGG 702 455 CACGACCTGGGTTTCCAGGA 703 456 ACGACCTGGGTTTCCAGGAG 704 457 CGACCTGGGTTTCCAGGAGG 705 458 GTCCCGGCGAGTACATCGTT 678 459 TGTCCCGGCGAGTACATCGT 677 460 CTGTCCCGGCGAGTACATCG 676 461 GCTGTCCCGGCGAGTACATC 675 462 CGCTGTCCCGGCGAGTACAT 674 463 TCGCTGTCCCGGCGAGTACA 673 464 ATCGCTGTCCCGGCGAGTAC 672 465 CATCGCTGTCCCGGCGAGTA 671 466 ACATCGCTGTCCCGGCGAGT 670 467 GACATCGCTGTCCCGGCGAG 669 468 AGACATCGCTGTCCCGGCGA 668 469 CAGACATCGCTGTCCCGGCG 667 470 GCAGACATCGCTGTCCCGGC 666 471 AGCAGACATCGCTGTCCCGG 665 472 CAGCAGACATCGCTGTCCCG 664 473 GCAGCAGACATCGCTGTCCC 663 474 TGCAGCAGACATCGCTGTCC 662 475 GTGCAGCAGACATCGCTGTC 661 476 AGTGCAGCAGACATCGCTGT 660 477 GAGTGCAGCAGACATCGCTG 659 478 GGAGTGCAGCAGACATCGCT 658 479 TGGAGTGCAGCAGACATCGC 657 480 ATGGAGTGCAGCAGACATCG 656 481 AACCTCCTCCCCGCCACGGGTT 1233 482 ACCTCCTCCCCGCCACGGGT 1234 483 CCTCCTCCCCGCCACGGGTT 1235 484 CTCCTCCCCGCCACGGGTTC 1236 485 TCCTCCCCGCCACGGGTTCA 1237 486 CCTCCCCGCCACGGGTTCAA 1238 487 CTCCCCGCCACGGGTTCAAG 1239 488 TCCCCGCCACGGGTTCAAGC 1240 489 CCCCGCCACGGGTTCAAGCG 1241 490 CCCGCCACGGGTTCAAGCGA 1242 491 CCGCCACGGGTTCAAGCGAT 1243 492 CGCCACGGGTTCAAGCGATT 1244 493 GCCACGGGTTCAAGCGATTC 1245 494 CCACGGGTTCAAGCGATTCT 1246 495 CACGGGTTCAAGCGATTCTC 1247 496 ACGGGTTCAAGCGATTCTCC 1248 497 CGGGTTCAAGCGATTCTCCT 1249 498 GGGTTCAAGCGATTCTCCTG 1250 499 GGTTCAAGCGATTCTCCTGC 1251 500 GTTCAAGCGATTCTCCTGCC 1252 501 TTCAAGCGATTCTCCTGCCT 1253 502 TCAAGCGATTCTCCTGCCTC 1254 503 CAAGCGATTCTCCTGCCTCA 1255 504 AAGCGATTCTCCTGCCTCAG 1256 505 AGCGATTCTCCTGCCTCAGC 1257 506 GCGATTCTCCTGCCTCAGCC 1258 507 CGATTCTCCTGCCTCAGCCT 1259 508 CAACCTCCTCCCCGCCACGG 1232 509 GCAACCTCCTCCCCGCCACG 1231 510 TGCAACCTCCTCCCCGCCAC 1230 511 CTGCAACCTCCTCCCCGCCA 1229 512 ACTGCAACCTCCTCCCCGCC 1228 513 CACTGCAACCTCCTCCCCGC 1227 514 TCACTGCAACCTCCTCCCCG 1226

Hot Zones (Relative upstream location to gene start site)  1-350 600-800 1100-1350 1900-2150 2750-3200

Examples

In FIG. 1, SU1 (1) shows a dose-dependent response in MDA-MB-231, a human breast cell line, with SU1 at 20 μL showing greater inhibition than SU1 at 10 and 3 μM. SU1's inhibition values, both at 20 and 10 μM, were statistically significant (P<0.05) compared to untreated control values. SU1's inhibition values at 3 μM were insignificant (insignificance indicated by bars with diagonal stripes). Furthermore, SU3's (3) inhibition values at 10 μM were insignificant compared to the untreated control values. SU3's diminished inhibition is attributable to the lack of a CG pair in the 5′ linear section before or at the base of the hairpin of the secondary structure and further back from the transcription start site compared to the other oligonucleotides tested. Two variants of SU1, SU1_(—)02 (4; 1 base shorter) and SU1_(—)03 (5; 4 bases longer), were also statistically significant at 10 μM (P<0.5) compared to the untreated control. This demonstrates that a sequence still retains its inhibitory levels despite shifting the sequence a few bases. The negative control (a scrambled oligonucleotide) was not statistically significant compared to the untreated control. The Survivin sequences SU1 (1), SU1_(—)02 (4), SU1_(—)03 (5) (shown below) fit the independent and dependent DNAi motif claims. As noted previously, SU3 (3), does not contain a CG in the 5′ linear section either prior to or in the base of the hairpin.

FIG. 2 is similar to FIG. 1 and in FIG. 2 it is shown that SU1 (1) demonstrated significant (P<0.05) inhibition of A549 (human lung cell line) compared to the untreated control values. Also, SU3's (3) inhibition values were insignificant compared to the untreated control values. The negative control was not statistically significant compared to the untreated control. The Survivin sequence SU1 (1) (shown below) fits the independent and dependent DNAi motif claims. As noted previously, SU3 (3), does not contain a CG in the 5′ linear section either prior to or in the base of the hairpin.

FIG. 3 shows that DU145 (human prostate cell line), SU1 (1) and its two variants, SU1_(—)02 (4) and SU1_(—)03 (5), produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated control values. SU2 (2), at 20 μM, produced statistically significant (P<0.05) inhibition compared to the untreated control values. The Survivin sequences SU1 (1), SU1_(—)02 (4), SU1_(—)03 (5), and SU2 (2) (shown below) fit the independent and dependent DNAi motif claims. As noted previously, SU3 (3), does not contain a CG in the 5′ linear section either prior to or in the base of the hairpin. SU2 (2) demonstrates that some oligonucleotides will show inhibition at acceptably higher concentrations (below a concentration where general cytotoxicity is observed) even though they may not demonstrate inhibition at lower concentrations.

FIG. 4 shows that in MCF7 (human mammary breast cell line), SU1 (1) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The Survivin sequence SU1 (1), fits the independent and dependent DNAi motif claims.

Secondary Structures FIGS. 5, 6, 7, 8, 9.

FIG. 5 is Sequence 1 (SU1). FIG. 6 is Sequence 2 (SU2). FIG. 7 is Sequence 3 (SU3) (Note in FIG. 7 or Sequence 3 there is No CG in the 5′ linear base. FIG. 8 is Sequence 4 (SU1_(—)02). FIG. 9 is Sequence 5 (SU1_(—)03).

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11950) ATCATGACACACATTTACCTGTGTAACAAACCTGCACATCCTACACA TATACCCTGGAACTTAAAGTAAAAGTTGGGGGGGGGGGTAAAAAAGAATT TCCACCGTGACATTATTGAGTATAGCAAAAAAAAAAAAAACAAGAAACAG CCTAGTGTTCATTAGGGAATAAACGCATTCAAGCAGCATCAAACCCTGCA GCCATTACAAAGAGATCTATGTTGACCATGTGGAATATCTCCAAGAGCCA CAGTAGCCTCCCTTATCTGTAGGATTCACTCCAAGACCCTCTGAAACCAT GGATAATACTGAACCCTATATACACTATGTTTTTTCTTGTATATACATAC CTACGATAAAGTTTAATTTATAAATTGGCAAAGGGTATATAAATATTCCT TCTAAGAGATTAACAATAACTAATAAAGTAGAACGATTAAAACAATATAC TGTGATCAAAGTTATGTGAAGCCAGGTGCTGTGGCTCATGCCTGTAATCC CAGCACTTTGGGAGGCTGAGACAGGTGGATCACCTGAGGTCAGGAGTTGG AGACCAGCCTGGCCAACATGACAAAACCCCGTCTCTACTAAAGATAAAAA AAATTAGCCGGGCATGGTGACACATGCCTGTAATCCCAGCTACTTGGGAG GCTGAGGCAGGAGAATCGCTTGAACCTGGGAGGCGGAGGTTGCAGTGAGC TAAGATCACACCATTGCACTCCAGCCTGGGCAACAAGAGTGAAACTCTGT CTCAAAACAAAACAAAACAAAACAAACTTATGGGGTTGCTCTCTTTCTCT CAAAATATCCTTTTTTTGGCAGGGCACGGTGGCTCATGCCTGTAATCCCA GCACTTTGAGAGGCTGAGGTGGGTGAATCACCTGAGGTCAGGAGTTCAAG ACCAGCCTGGCCAACATGGTGAAACCCCGTCTCTATTAAAAATACAAAAA ATTAGCTGGGCGTGGTGGTGCAGGCCTGTAATCCCAGCTACTTGGGAGGC TGAGGCAGGAGAATCACTCGAACCCAGGAGCTGGAGTTTGCAGTGAGCCG AGATCATGCCATTGCACTCCAGCCTGGGCCACAGAGCAAGACTCCATCTC AAAAAAAAAAAAAAGAAAAAAAGAAAGTCTTTTTTTTTTTTGAGACTGTA TCTCACTCTTTCTCCCAGGCTGGAGTGCAGTGGCCCAATCATGGCTCACT GCAGCCTCGACCTCCCAGGATCAAGTGATCCTTCCACCTCAGCCTCCCGA GTAGCTGGAAGTATAGGTGCACGCCCGACTGATTTTTTTTTTTTTTTTTA GACGGAGTCTCACTCTTGTTGCTCTGGCTGGAGTGCAATGGCAGGATCTC GGCTCACTGCAACCTCTGCCTCTTAGATTCAAGCGATTCTCGTGCCTCAG CCTCCCGAGTAGCTGGGATTACAGGTGCCCACCACCATGCCCGGATAATT TTTTGTATTTTTAATAGAGACAGGGTTTCACCATATTGGTCAGGCTGGTC TCAAACTCCTGACCTCAGGTGATCCACCTGCCTCAGCCTCCCAAACTGCT GGGATTACAGGCGTGAGCCACCGGGCATGGCCTTTCCTGGCTAATTTTTT AAATTTTTGATAGAGATGGGGTCTCAGTGTTGCCCAGGCTGATCTTGAAC TCCTAGATTCAAGTGATCCTCCCTCCTTGGTCTCCCAAAGTGCTGAGATT ACAGGCGTGAGCCACCGCCCCGGGCTGGAAAATACTTTTTTAAACGAGGG CAATGTGAATCTGAAATGCCATTTGAGGAAAGATCTGTTCGCCTGACATC CTGTTTGAGCCTGGGTGGACAGGACAGCACCTGCCAGCATCGGGAAGCAC TGCAGATGGGAAGAGGCTTGGTCACTCTCCAAAGGTGGCAGGAGTTGGAG GGGGTGAGCTGAAGGTAAGGAGAAAGGAGGTGGGGACCCAGGAGACAGGG GCTGCGCAGCGGGCTCGGGGCTGACACCCCCACGGATACAGTTCACTGGG GCTCAAACATAAAAGGAACCCAACTATTGTGGGAGGAAAAGACTCTTCTG CCTTTCTGCCTTTTCTTTTTTTCTTTTTCTTTCTTTCTTTTTTTTTTTTT TTTTTTGAGACAGAGTCTTGCTCTATCGCCCAGGCTGGAGTGCAGTGGCG TGATCTCGGCTCACTGCAAGCTCTGCCTCCCGGGATCACGCCATTCTCCT GCCTCAACCTCCCGAGCAGCTGGGACTACAGGCGCCTGCCACCACACCCG GCTATTTTTTTGTATTTTTTAGTAGAGATGGGGTTTCACCGTGTTAGCCA GGACGGTCTCGATCTCCTGACCTTGTGATCCGCCCGCCTCGGCCTCCCAA AGTGCTGGGATTACAGGCGTGAGCCACCGCGCCTGGCTCTTTTTTCTTTC TTTTTTTTTTTTCCGAGACAGAGTTTCACTCTTGTTGCCCAGGCTGGAGT GCAGTGGCGCAATCTTGGCTCACTGCAACCTCCACCTCCAGGGTTCAAGC GATTCTCCTGCCTCAGCCTCCTGAGTAGCTGGGACTGCAGGCGCGCACCA CCACGCCTGGCTAATTTTTGTATTTTTAGTAGAGACAGGGTTTCACCATA TTGGCCAGGCTGGTCTCGAACTCCTGACCTTGTGATCTGCCCACCTCAGC CTCCCAAAGTCCTGGGATTACAGGCGTGAGCCACCGTGCCCAGCCTGACC CCTCTGCCCTTTCAAAAACTATGTTCGTTCTCTCACAGCCTTCTCTTGTC ATATTAAGTCCACACCGCAGGCCTAATTTGTCCAGTGAATGCTATGCAAA TATTTCATGCACCTGCTGATCGCAGGAATGATATGTACTTGGTACGCACT GATCGTACCTCGGGGTGGGAGAAGAGAGGGCAAGGAAGCAAAGAATAGCC CCCTCCTTTCCTGGTGCACCTTCAGATGTGCCGATGGGGCCCAGGCTCGC TGCAGATGGCCCCCTTCCCAGAGACAGGGGAGGATCCTCCACCCACTCCC CAGCCTCCAGGACCATCCTGACTCCTGCCTTCAGGCACTCAAGTTATGCG TCTAGACATGCGGATATATTCAAGCTGGGCACAGCACAGCAGCCCCACCC CAGGCAGCTTGAAATCAGAGCTGGGGTCCAAAGGGACCACACCCCGAGGG ACTGTGTGGGGGTCGGGGCACACAGGCCACTGCTTCCCCCCGTCTTTCTC AGCCATTCCTGAAGTCAGCCTCACTCTGCTTCTCAGGGATTTCAAATGTG CAGAGACTCTGGCACTTTTGTAGAAGCCCCTTCTGGTCCTAACTTACACC TGGATGCTGTGGGGCTGCAGCTGCTGCTCGGGCTCGGGAGGATGCTGGGG GCCCGGTGCCCATGAGCTTTTGAAGCTCCTGGAACTCGGTTTTGAGGGTG TTCAGGTCCAGGTGGACACCTGGGCTGTCCTTGTCCATGCATTTGATGAC ATTGTGTGCAGAAGTGAAAAGGAGTTAGGCCGGGCATGCTGGCTTATGCC TGTAATCCCAGCACTTTGGGAGGCTGAGGCGGGTGGATCACGAGGTCAGG AGTTCAATACCAGCCTGGCCAAGATGGTGAAACCCCGTCTCTACTAAAAA TACAAAAAAATTAGCCGGGCATGGTGGCGGGCGCATGTAATCCCAGCTAC TGGGGGGGCTGAGGCAGAGAATTGCTGGAACCCAGGAGATGGAGGTTGCA GTGAGCCAAGATTGTGCCACTGCACTGCACTCCAGCCTGGCGACAGAGCA AGACTCTGTCTCAAAAAAAAAAAAAAAAAGTGAAAAGGAGTTGTTCCTTT CCTCCCTCCTGAGGGCAGGCAACTGCTGCGGTTGCCAGTGGAGGTGGTGC GTCCTTGGTCTGTGCCTGGGGGCCACCCCAGCAGAGGCCATGGTGGTGCC AGGGCCCGGTTAGCGAGCCAATCAGCAGGACCCAGGGGCGACCTGCCAAA GTCAACTGGATTTGATAACTGCAGCGAAGTTAAGTTTCCTGATTTTGATG ATTGTGTTGTGGTTGTGTAAGAGAATGAAGTATTTCGGGGTAGTATGGTA ATGCCTTCAACTTACAAACGGTTCAGGTAAACCACCCATATACATACATA TACATGCATGTGATATATACACATACAGGGATGTGTGTGTGTTCACATAT ATGAGGGGAGAGAGACTAGGGGAGAGAAAGTAGGTTGGGGAGAGGGAGAG AGAAAGGAAAACAGGAGACAGAGAGAGAGCGGGGAGTAGAGAGAGGGAAG GGGTAAGAGAGGGAGAGGAGGAGAGAAAGGGAGGAAGAAGCAGAGAGTGA ATGTTAAAGGAAACAGGCAAAACATAAACAGAAAATCTGGGTGAAGGGTA TATGAGTATTCTTTGTACTATTCTTGCAATTATCTTTTATTTAAATTGAC ATCGGGCCGGGCGCAGTGGCTCACATCTGTAATCCCAGCACTTTGGGAGG CCGAGGCAGGCAGATCACTTGAGGTCAGGAGTTTGAGACCAGCCTGGCAA ACATGGTGAAACCCCATCTCTACTAAAAATACAAAAATTAGCCTGGTGTG GTGGTGCATGCCTTTAATCTCAGCTACTCGGGAGGCTGAGGCAGGAGAAT CGCTTGAACCCGTGGCGGGGAGGAGGTTGCAGTGAGCTGAGATCATGCCA CTGCACTCCAGCCTGGGCGATAGAGCGAGACTCAGTTTCAAATAAATAAA TAAACATCAAAATAAAAAGTTACTGTATTAAAGAATGGGGGCGGGGTGGG AGGGGTGGGGAGAGGTTGCAAAAATAAATAAATAAATAAATAAACCCCAA AATGAAAAAGACAGTGGAGGCACCAGGCCTGCGTGGGGCTGGAGGGCTAA TAAGGCCAGGCCTCTTATCTCTGGCCATAGAACCAGAGAAGTGAGTGGAT GTGATGCCCAGCTCCAGAAGTGACTCCAGAACACCCTGTTCCAAAGCAGA GGACACACTGATTTTTTTTTTAATAGGCTGCAGGACTTACTGTTGGTGGG ACGCCCTGCTTTGCGAAGGGAAAGGAGGAGTTTGCCCTGAGCACAGGCCC CCACCCTCCACTGGGCTTTCCCCAGCTCCCTTGTCTTCTTATCACGGTAG TGGCCCAGTCCCTGGCCCCTGACTCCAGAAGGTGGCCCTCCTGGAAACCC AGGTCGTGCAGTCAACGATGTACTCGCCGGGACAGCGATGTCTGCTGCAC TCCATCCCTCCCCTGTTCATTTGTCCTTCATGCCCGTCTGGAGTAGATGC TTTTTGCAGAGGTGGCACCCTGTAAAGCTCTCCTGTCTGACTTTTTTTTT TTTTTTAGACTGAGTTTTGCTCTTGTTGCCTAGGCTGGAGTGCAATGGCA CAATCTCAGCTCACTGCACCCTCTGCCTCCCGGGTTCAAGCGATTCTCCT GCCTCAGCCTCCCGAGTAGTTGGGATTACAGGCATGCACCACCACGCCCA GCTAATTTTTGTATTTTTAGTAGAGACAAGGTTTCACCGTGATGGCCAGG CTGGTCTTGAACTCCAGGACTCAAGTGATGCTCCTGCCTAGGCCTCTCAA AGTGTTGGGATTACAGGCGTGAGCCACTGCACCCGGCCTGCACGCGTTCT TTGAAAGCAGTCGAGGGGGCGCTAGGTGTGGGCAGGGACGAGCTGGCGCG GCGTCGCTGGGTGCACCGCGACCACGGGCAGAGCCACGCGGCGGGAGGAC TACAACTCCCGGCACACCCCGCGCCGCCCCGCCTCTACTCCCAGAAGGCC GCGGGGGGTGGACCGCCTAAGAGGGCGTGCGCTCCCGACATGCCCCGCGG CGCGCCATTAACCGCCAGATTTGAATCGCGGGACCCGTTGGCAGAGGTGG CGGCGGCGGC ATG

2) Beclin-1. Beclin 1, the mammalian orthologue of yeast Atg6, has a central role in autophagy, a process of programmed cell survival, which is increased during periods of cell stress and extinguished during the cell cycle. It interacts with several cofactors (Atg14L, UVRAG, Bif-1, Rubicon, Ambra1, HMGB1, nPIST, VMP1, SLAM, IP₃R, PINK and survivin) to regulate the lipid kinase Vps-34 protein and promote formation of Beclin 1-Vps34-Vps15 core complexes, thereby inducing autophagy. In contrast, the BH3 domain of Beclin 1 is bound to, and inhibited by Bcl-2 or Bcl-XL. This interaction can be disrupted by phosphorylation of Bcl-2 and Beclin 1, or ubiquitination of Beclin 1. Interestingly, caspase-mediated cleavage of Beclin 1 promotes crosstalk between apoptosis and autophagy. Beclin 1 dysfunction has been implicated in many disorders, including cancer and neurodegeneration (reviewed by Kang et al., Cell Death Differ. 2011 April; 18(4): 571-580).

Protein: Beclin-1 Gene: BECN1 (Homo sapiens, chromosome 17, 40962150-40976310 [NCBI Reference Sequence: NC_(—)000017.10]; start site location: 40975895; strand: negative)

Gene Identification GeneID 8678 HGNC 1034 HPRD 05087 MIM 604378

Targeted Sequences Relative upstream location Se- De- to gene quence sign start ID No: ID Sequence (5′-3′) site 515 BE1 CGACGCCCTTGACCTCCGGCCCGGGGT 39 550 BE2 CTGCGCCGTTCCCTCTAGGAATGG 111 572 GAAGCGACGCCCTTGACCTCCGGCCCGG 35 607 CCCCCGATGCTCTTCACCTCGGG 261 712 CGGGTCGGCCCCGGAGCGAGGCC 335 817 GCCCGGCAGCGGCCCCCAGAGGCCG 475 847 CGGTCTACCGCGGAGGCACTGTGGCCTCGG 308 952 ACAAAAACTAGCCGGGCGTGGTGGGGCACG 735 CC

Target Shift Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 515 CGACGCCCTTGACCTCCGGCCCGGGGT 39 516 GACGCCCTTGACCTCCGGCC 40 517 ACGCCCTTGACCTCCGGCCC 41 518 CGCCCTTGACCTCCGGCCCG 42 519 GCCCTTGACCTCCGGCCCGG 43 520 CCCTTGACCTCCGGCCCGGG 44 521 CCTTGACCTCCGGCCCGGGG 45 522 CTTGACCTCCGGCCCGGGGT 46 523 TTGACCTCCGGCCCGGGGTT 47 524 TGACCTCCGGCCCGGGGTTA 48 525 GACCTCCGGCCCGGGGTTAC 49 526 ACCTCCGGCCCGGGGTTACC 50 527 CCTCCGGCCCGGGGTTACCA 51 528 CTCCGGCCCGGGGTTACCAC 52 529 TCCGGCCCGGGGTTACCACA 53 530 CCGGCCCGGGGTTACCACAT 54 531 CGGCCCGGGGTTACCACATG 55 532 GGCCCGGGGTTACCACATGC 56 533 GCCCGGGGTTACCACATGCC 57 534 CCCGGGGTTACCACATGCCT 58 535 CCGGGGTTACCACATGCCTT 59 536 CGGGGTTACCACATGCCTTG 60 537 GCGACGCCCTTGACCTCCGG 38 538 AGCGACGCCCTTGACCTCCG 37 539 AAGCGACGCCCTTGACCTCC 36 540 GAAGCGACGCCCTTGACCTC 35 541 AGAAGCGACGCCCTTGACCT 34 542 GAGAAGCGACGCCCTTGACC 33 543 GGAGAAGCGACGCCCTTGAC 32 544 GGGAGAAGCGACGCCCTTGA 31 545 AGGGAGAAGCGACGCCCTTG 30 546 TAGGGAGAAGCGACGCCCTT 29 547 TTAGGGAGAAGCGACGCCCT 28 548 ATTAGGGAGAAGCGACGCCC 27 549 CATTAGGGAGAAGCGACGCC 26 550 CTGCGCCGTTCCCTCTAGGAATGG 111 551 TGCGCCGTTCCCTCTAGGAA 112 552 GCGCCGTTCCCTCTAGGAAT 113 553 CGCCGTTCCCTCTAGGAATG 114 554 GCCGTTCCCTCTAGGAATGG 115 555 CCGTTCCCTCTAGGAATGGT 116 556 CGTTCCCTCTAGGAATGGTA 117 557 CCTGCGCCGTTCCCTCTAGG 110 558 ACCTGCGCCGTTCCCTCTAG 109 559 AACCTGCGCCGTTCCCTCTA 108 560 CAACCTGCGCCGTTCCCTCT 107 561 CCAACCTGCGCCGTTCCCTC 106 562 CCCAACCTGCGCCGTTCCCT 105 563 TCCCAACCTGCGCCGTTCCC 104 564 GTCCCAACCTGCGCCGTTCC 103 565 AGTCCCAACCTGCGCCGTTC 102 566 AAGTCCCAACCTGCGCCGTT 101 567 GAAGTCCCAACCTGCGCCGT 100 568 GGAAGTCCCAACCTGCGCCG 99 569 GGGAAGTCCCAACCTGCGCC 98 570 AGGGAAGTCCCAACCTGCGC 97 571 GAGGGAAGTCCCAACCTGCG 96 572 GAAGCGACGCCCTTGACCTCCGGCCCGG 35 573 AAGCGACGCCCTTGACCTCC 36 574 AGCGACGCCCTTGACCTCCG 37 575 GCGACGCCCTTGACCTCCGG 38 576 CGACGCCCTTGACCTCCGGC 39 577 GACGCCCTTGACCTCCGGCC 40 578 ACGCCCTTGACCTCCGGCCC 41 579 CGCCCTTGACCTCCGGCCCG 42 580 GCCCTTGACCTCCGGCCCGG 43 581 CCCTTGACCTCCGGCCCGGG 44 582 CCTTGACCTCCGGCCCGGGG 45 583 CTTGACCTCCGGCCCGGGGT 46 584 TTGACCTCCGGCCCGGGGTT 47 585 TGACCTCCGGCCCGGGGTTA 48 586 GACCTCCGGCCCGGGGTTAC 49 587 ACCTCCGGCCCGGGGTTACC 50 588 CCTCCGGCCCGGGGTTACCA 51 589 CTCCGGCCCGGGGTTACCAC 52 590 TCCGGCCCGGGGTTACCACA 53 591 CCGGCCCGGGGTTACCACAT 54 592 CGGCCCGGGGTTACCACATG 55 593 GGCCCGGGGTTACCACATGC 56 594 GCCCGGGGTTACCACATGCC 57 595 CCCGGGGTTACCACATGCCT 58 596 CCGGGGTTACCACATGCCTT 59 597 CGGGGTTACCACATGCCTTG 60 598 AGAAGCGACGCCCTTGACCT 34 599 GAGAAGCGACGCCCTTGACC 33 600 GGAGAAGCGACGCCCTTGAC 32 601 GGGAGAAGCGACGCCCTTGA 31 602 AGGGAGAAGCGACGCCCTTG 30 603 TAGGGAGAAGCGACGCCCTT 29 604 TTAGGGAGAAGCGACGCCCT 28 605 ATTAGGGAGAAGCGACGCCC 27 606 CATTAGGGAGAAGCGACGCC 26 607 CCCCCGATGCTCTTCACCTCGGG 261 608 CCCCGATGCTCTTCACCTCG 262 609 CCCGATGCTCTTCACCTCGG 263 610 CCGATGCTCTTCACCTCGGG 264 611 CGATGCTCTTCACCTCGGGA 265 612 GATGCTCTTCACCTCGGGAG 266 613 ATGCTCTTCACCTCGGGAGC 267 614 TGCTCTTCACCTCGGGAGCC 268 615 GCTCTTCACCTCGGGAGCCC 269 616 CTCTTCACCTCGGGAGCCCG 270 617 TCTTCACCTCGGGAGCCCGG 271 618 CTTCACCTCGGGAGCCCGGA 272 619 TTCACCTCGGGAGCCCGGAG 273 620 TCACCTCGGGAGCCCGGAGC 274 621 CACCTCGGGAGCCCGGAGCC 275 622 ACCTCGGGAGCCCGGAGCCC 276 623 CCTCGGGAGCCCGGAGCCCG 277 624 CTCGGGAGCCCGGAGCCCGT 278 625 TCGGGAGCCCGGAGCCCGTC 279 626 CGGGAGCCCGGAGCCCGTCA 280 627 GGGAGCCCGGAGCCCGTCAC 281 628 GGAGCCCGGAGCCCGTCACC 282 629 GAGCCCGGAGCCCGTCACCC 283 630 AGCCCGGAGCCCGTCACCCA 284 631 GCCCGGAGCCCGTCACCCAA 285 632 CCCGGAGCCCGTCACCCAAG 286 633 CCGGAGCCCGTCACCCAAGT 287 634 CGGAGCCCGTCACCCAAGTC 288 635 GGAGCCCGTCACCCAAGTCC 289 636 GAGCCCGTCACCCAAGTCCG 290 637 AGCCCGTCACCCAAGTCCGG 291 638 GCCCGTCACCCAAGTCCGGT 292 639 CCCGTCACCCAAGTCCGGTC 293 640 CCGTCACCCAAGTCCGGTCT 294 641 CGTCACCCAAGTCCGGTCTA 295 642 GTCACCCAAGTCCGGTCTAC 296 643 TCACCCAAGTCCGGTCTACC 297 644 CACCCAAGTCCGGTCTACCG 298 645 ACCCAAGTCCGGTCTACCGC 299 646 CCCAAGTCCGGTCTACCGCG 300 647 CCAAGTCCGGTCTACCGCGG 301 648 CAAGTCCGGTCTACCGCGGA 302 649 AAGTCCGGTCTACCGCGGAG 303 650 AGTCCGGTCTACCGCGGAGG 304 651 GTCCGGTCTACCGCGGAGGC 305 652 TCCGGTCTACCGCGGAGGCA 306 653 CCGGTCTACCGCGGAGGCAC 307 654 CGGTCTACCGCGGAGGCACT 308 655 GGTCTACCGCGGAGGCACTG 309 656 GTCTACCGCGGAGGCACTGT 310 657 TCTACCGCGGAGGCACTGTG 311 658 CTACCGCGGAGGCACTGTGG 312 659 TACCGCGGAGGCACTGTGGC 313 660 ACCGCGGAGGCACTGTGGCC 314 661 CCGCGGAGGCACTGTGGCCT 315 662 CGCGGAGGCACTGTGGCCTC 316 663 GCGGAGGCACTGTGGCCTCG 317 664 CGGAGGCACTGTGGCCTCGG 318 665 GGAGGCACTGTGGCCTCGGG 319 666 GAGGCACTGTGGCCTCGGGT 320 667 AGGCACTGTGGCCTCGGGTC 321 668 GGCACTGTGGCCTCGGGTCG 322 669 GCACTGTGGCCTCGGGTCGG 323 670 CACTGTGGCCTCGGGTCGGC 324 671 ACTGTGGCCTCGGGTCGGCC 325 672 CTGTGGCCTCGGGTCGGCCC 326 673 TGTGGCCTCGGGTCGGCCCC 327 674 GTGGCCTCGGGTCGGCCCCG 328 675 TGGCCTCGGGTCGGCCCCGG 329 676 GGCCTCGGGTCGGCCCCGGA 330 677 GCCTCGGGTCGGCCCCGGAG 331 678 CCTCGGGTCGGCCCCGGAGC 332 679 CTCGGGTCGGCCCCGGAGCG 333 680 TCGGGTCGGCCCCGGAGCGA 334 681 CGGGTCGGCCCCGGAGCGAG 335 682 GGGTCGGCCCCGGAGCGAGG 336 683 GGTCGGCCCCGGAGCGAGGC 337 684 GTCGGCCCCGGAGCGAGGCC 338 685 TCGGCCCCGGAGCGAGGCCT 339 686 CGGCCCCGGAGCGAGGCCTC 340 687 GGCCCCGGAGCGAGGCCTCC 341 688 GCCCCGGAGCGAGGCCTCCA 342 689 CCCCGGAGCGAGGCCTCCAG 343 690 CCCGGAGCGAGGCCTCCAGA 344 691 CCGGAGCGAGGCCTCCAGAA 345 692 CGGAGCGAGGCCTCCAGAAC 346 693 GGAGCGAGGCCTCCAGAACT 347 694 GAGCGAGGCCTCCAGAACTA 348 695 AGCGAGGCCTCCAGAACTAC 349 696 GCGAGGCCTCCAGAACTACC 350 697 CGAGGCCTCCAGAACTACCA 351 698 GCCCCCGATGCTCTTCACCT 260 699 AGCCCCCGATGCTCTTCACC 259 700 CAGCCCCCGATGCTCTTCAC 258 701 TCAGCCCCCGATGCTCTTCA 257 702 CTCAGCCCCCGATGCTCTTC 256 703 CCTCAGCCCCCGATGCTCTT 255 704 ACCTCAGCCCCCGATGCTCT 254 705 CACCTCAGCCCCCGATGCTC 253 706 CCACCTCAGCCCCCGATGCT 252 707 CCCACCTCAGCCCCCGATGC 251 708 TCCCACCTCAGCCCCCGATG 250 709 GTCCCACCTCAGCCCCCGAT 249 710 GGTCCCACCTCAGCCCCCGA 248 711 AGGTCCCACCTCAGCCCCCG 247 712 CGGGTCGGCCCCGGAGCGAGGCC 335 713 GGGTCGGCCCCGGAGCGAGG 336 714 GGTCGGCCCCGGAGCGAGGC 337 715 GTCGGCCCCGGAGCGAGGCC 338 716 TCGGCCCCGGAGCGAGGCCT 339 717 CGGCCCCGGAGCGAGGCCTC 340 718 GGCCCCGGAGCGAGGCCTCC 341 719 GCCCCGGAGCGAGGCCTCCA 342 720 CCCCGGAGCGAGGCCTCCAG 343 721 CCCGGAGCGAGGCCTCCAGA 344 722 CCGGAGCGAGGCCTCCAGAA 345 723 CGGAGCGAGGCCTCCAGAAC 346 724 GGAGCGAGGCCTCCAGAACT 347 725 GAGCGAGGCCTCCAGAACTA 348 726 AGCGAGGCCTCCAGAACTAC 349 727 GCGAGGCCTCCAGAACTACC 350 728 CGAGGCCTCCAGAACTACCA 351 729 TCGGGTCGGCCCCGGAGCGA 334 730 CTCGGGTCGGCCCCGGAGCG 333 731 CCTCGGGTCGGCCCCGGAGC 332 732 GCCTCGGGTCGGCCCCGGAG 331 733 GGCCTCGGGTCGGCCCCGGA 330 734 TGGCCTCGGGTCGGCCCCGG 329 735 GTGGCCTCGGGTCGGCCCCG 328 736 TGTGGCCTCGGGTCGGCCCC 327 737 CTGTGGCCTCGGGTCGGCCC 326 738 ACTGTGGCCTCGGGTCGGCC 325 739 CACTGTGGCCTCGGGTCGGC 324 740 GCACTGTGGCCTCGGGTCGG 323 741 GGCACTGTGGCCTCGGGTCG 322 742 AGGCACTGTGGCCTCGGGTC 321 743 GAGGCACTGTGGCCTCGGGT 320 744 GGAGGCACTGTGGCCTCGGG 319 745 CGGAGGCACTGTGGCCTCGG 318 746 GCGGAGGCACTGTGGCCTCG 317 747 CGCGGAGGCACTGTGGCCTC 316 748 CCGCGGAGGCACTGTGGCCT 315 749 ACCGCGGAGGCACTGTGGCC 314 750 TACCGCGGAGGCACTGTGGC 313 751 CTACCGCGGAGGCACTGTGG 312 752 TCTACCGCGGAGGCACTGTG 311 753 GTCTACCGCGGAGGCACTGT 310 754 GGTCTACCGCGGAGGCACTG 309 755 CGGTCTACCGCGGAGGCACT 308 756 CCGGTCTACCGCGGAGGCAC 307 757 TCCGGTCTACCGCGGAGGCA 306 758 GTCCGGTCTACCGCGGAGGC 305 759 AGTCCGGTCTACCGCGGAGG 304 760 AAGTCCGGTCTACCGCGGAG 303 761 CAAGTCCGGTCTACCGCGGA 302 762 CCAAGTCCGGTCTACCGCGG 301 763 CCCAAGTCCGGTCTACCGCG 300 764 ACCCAAGTCCGGTCTACCGC 299 765 CACCCAAGTCCGGTCTACCG 298 766 TCACCCAAGTCCGGTCTACC 297 767 GTCACCCAAGTCCGGTCTAC 296 768 CGTCACCCAAGTCCGGTCTA 295 769 CCGTCACCCAAGTCCGGTCT 294 770 CCCGTCACCCAAGTCCGGTC 293 771 GCCCGTCACCCAAGTCCGGT 292 772 AGCCCGTCACCCAAGTCCGG 291 773 GAGCCCGTCACCCAAGTCCG 290 774 GGAGCCCGTCACCCAAGTCC 289 775 CGGAGCCCGTCACCCAAGTC 288 776 CCGGAGCCCGTCACCCAAGT 287 777 CCCGGAGCCCGTCACCCAAG 286 778 GCCCGGAGCCCGTCACCCAA 285 779 AGCCCGGAGCCCGTCACCCA 284 780 GAGCCCGGAGCCCGTCACCC 283 781 GGAGCCCGGAGCCCGTCACC 282 782 GGGAGCCCGGAGCCCGTCAC 281 783 CGGGAGCCCGGAGCCCGTCA 280 784 TCGGGAGCCCGGAGCCCGTC 279 785 CTCGGGAGCCCGGAGCCCGT 278 786 CCTCGGGAGCCCGGAGCCCG 277 787 ACCTCGGGAGCCCGGAGCCC 276 788 CACCTCGGGAGCCCGGAGCC 275 789 TCACCTCGGGAGCCCGGAGC 274 790 TTCACCTCGGGAGCCCGGAG 273 791 CTTCACCTCGGGAGCCCGGA 272 792 TCTTCACCTCGGGAGCCCGG 271 793 CTCTTCACCTCGGGAGCCCG 270 794 GCTCTTCACCTCGGGAGCCC 269 795 TGCTCTTCACCTCGGGAGCC 268 796 ATGCTCTTCACCTCGGGAGC 267 797 GATGCTCTTCACCTCGGGAG 266 798 CGATGCTCTTCACCTCGGGA 265 799 CCGATGCTCTTCACCTCGGG 264 800 CCCGATGCTCTTCACCTCGG 263 801 CCCCGATGCTCTTCACCTCG 262 802 CCCCCGATGCTCTTCACCTC 261 803 GCCCCCGATGCTCTTCACCT 260 804 AGCCCCCGATGCTCTTCACC 259 805 CAGCCCCCGATGCTCTTCAC 258 806 TCAGCCCCCGATGCTCTTCA 257 807 CTCAGCCCCCGATGCTCTTC 256 808 CCTCAGCCCCCGATGCTCTT 255 809 ACCTCAGCCCCCGATGCTCT 254 810 CACCTCAGCCCCCGATGCTC 253 811 CCACCTCAGCCCCCGATGCT 252 812 CCCACCTCAGCCCCCGATGC 251 813 TCCCACCTCAGCCCCCGATG 250 814 GTCCCACCTCAGCCCCCGAT 249 815 GGTCCCACCTCAGCCCCCGA 248 816 AGGTCCCACCTCAGCCCCCG 247 817 GCCCGGCAGCGGCCCCCAGAGGCCG 475 818 CCCGGCAGCGGCCCCCAGAG 476 819 CCGGCAGCGGCCCCCAGAGG 477 820 CGGCAGCGGCCCCCAGAGGC 478 821 GGCAGCGGCCCCCAGAGGCC 479 822 GCAGCGGCCCCCAGAGGCCG 480 823 CAGCGGCCCCCAGAGGCCGG 481 824 AGCGGCCCCCAGAGGCCGGG 482 825 GCGGCCCCCAGAGGCCGGGC 483 826 CGGCCCCCAGAGGCCGGGCT 484 827 GGCCCCCAGAGGCCGGGCTG 485 828 GCCCCCAGAGGCCGGGCTGG 486 829 CCCCCAGAGGCCGGGCTGGG 487 830 CCCCAGAGGCCGGGCTGGGA 488 831 CCCAGAGGCCGGGCTGGGAA 489 832 GGCCCGGCAGCGGCCCCCAG 474 833 AGGCCCGGCAGCGGCCCCCA 473 834 CAGGCCCGGCAGCGGCCCCC 472 835 ACAGGCCCGGCAGCGGCCCC 471 836 CACAGGCCCGGCAGCGGCCC 470 837 TCACAGGCCCGGCAGCGGCC 469 838 CTCACAGGCCCGGCAGCGGC 468 839 GCTCACAGGCCCGGCAGCGG 467 840 GGCTCACAGGCCCGGCAGCG 466 841 AGGCTCACAGGCCCGGCAGC 465 842 CAGGCTCACAGGCCCGGCAG 464 843 ACAGGCTCACAGGCCCGGCA 463 844 CACAGGCTCACAGGCCCGGC 462 845 CCACAGGCTCACAGGCCCGG 461 846 TCCACAGGCTCACAGGCCCG 460 847 CGGTCTACCGCGGAGGCACTGTGGCCTCGG 308 848 GGTCTACCGCGGAGGCACTG 309 849 GTCTACCGCGGAGGCACTGT 310 850 TCTACCGCGGAGGCACTGTG 311 851 CTACCGCGGAGGCACTGTGG 312 852 TACCGCGGAGGCACTGTGGC 313 853 ACCGCGGAGGCACTGTGGCC 314 854 CCGCGGAGGCACTGTGGCCT 315 855 CGCGGAGGCACTGTGGCCTC 316 856 GCGGAGGCACTGTGGCCTCG 317 857 CGGAGGCACTGTGGCCTCGG 318 858 GGAGGCACTGTGGCCTCGGG 319 859 GAGGCACTGTGGCCTCGGGT 320 860 AGGCACTGTGGCCTCGGGTC 321 861 GGCACTGTGGCCTCGGGTCG 322 862 GCACTGTGGCCTCGGGTCGG 323 863 CACTGTGGCCTCGGGTCGGC 324 864 ACTGTGGCCTCGGGTCGGCC 325 865 CTGTGGCCTCGGGTCGGCCC 326 866 TGTGGCCTCGGGTCGGCCCC 327 867 GTGGCCTCGGGTCGGCCCCG 328 868 TGGCCTCGGGTCGGCCCCGG 329 869 GGCCTCGGGTCGGCCCCGGA 330 870 GCCTCGGGTCGGCCCCGGAG 331 871 CCTCGGGTCGGCCCCGGAGC 332 872 CTCGGGTCGGCCCCGGAGCG 333 873 TCGGGTCGGCCCCGGAGCGA 334 874 CGGGTCGGCCCCGGAGCGAG 335 875 GGGTCGGCCCCGGAGCGAGG 336 876 GGTCGGCCCCGGAGCGAGGC 337 877 GTCGGCCCCGGAGCGAGGCC 338 878 TCGGCCCCGGAGCGAGGCCT 339 879 CGGCCCCGGAGCGAGGCCTC 340 880 GGCCCCGGAGCGAGGCCTCC 341 881 GCCCCGGAGCGAGGCCTCCA 342 882 CCCCGGAGCGAGGCCTCCAG 343 883 CCCGGAGCGAGGCCTCCAGA 344 884 CCGGAGCGAGGCCTCCAGAA 345 885 CGGAGCGAGGCCTCCAGAAC 346 886 GGAGCGAGGCCTCCAGAACT 347 887 GAGCGAGGCCTCCAGAACTA 348 888 AGCGAGGCCTCCAGAACTAC 349 889 GCGAGGCCTCCAGAACTACC 350 890 CGAGGCCTCCAGAACTACCA 351 891 CCGGTCTACCGCGGAGGCAC 307 892 TCCGGTCTACCGCGGAGGCA 306 893 GTCCGGTCTACCGCGGAGGC 305 894 AGTCCGGTCTACCGCGGAGG 304 895 AAGTCCGGTCTACCGCGGAG 303 896 CAAGTCCGGTCTACCGCGGA 302 897 CCAAGTCCGGTCTACCGCGG 301 898 CCCAAGTCCGGTCTACCGCG 300 899 ACCCAAGTCCGGTCTACCGC 299 900 CACCCAAGTCCGGTCTACCG 298 901 TCACCCAAGTCCGGTCTACC 297 902 GTCACCCAAGTCCGGTCTAC 296 903 CGTCACCCAAGTCCGGTCTA 295 904 CCGTCACCCAAGTCCGGTCT 294 905 CCCGTCACCCAAGTCCGGTC 293 906 GCCCGTCACCCAAGTCCGGT 292 907 AGCCCGTCACCCAAGTCCGG 291 908 GAGCCCGTCACCCAAGTCCG 290 909 GGAGCCCGTCACCCAAGTCC 289 910 CGGAGCCCGTCACCCAAGTC 288 911 CCGGAGCCCGTCACCCAAGT 287 912 CCCGGAGCCCGTCACCCAAG 286 913 GCCCGGAGCCCGTCACCCAA 285 914 AGCCCGGAGCCCGTCACCCA 284 915 GAGCCCGGAGCCCGTCACCC 283 916 GGAGCCCGGAGCCCGTCACC 282 917 GGGAGCCCGGAGCCCGTCAC 281 918 CGGGAGCCCGGAGCCCGTCA 280 919 TCGGGAGCCCGGAGCCCGTC 279 920 CTCGGGAGCCCGGAGCCCGT 278 921 CCTCGGGAGCCCGGAGCCCG 277 922 ACCTCGGGAGCCCGGAGCCC 276 923 CACCTCGGGAGCCCGGAGCC 275 924 TCACCTCGGGAGCCCGGAGC 274 925 TTCACCTCGGGAGCCCGGAG 273 926 CTTCACCTCGGGAGCCCGGA 272 927 TCTTCACCTCGGGAGCCCGG 271 928 CTCTTCACCTCGGGAGCCCG 270 929 GCTCTTCACCTCGGGAGCCC 269 930 TGCTCTTCACCTCGGGAGCC 268 931 ATGCTCTTCACCTCGGGAGC 267 932 GATGCTCTTCACCTCGGGAG 266 933 CGATGCTCTTCACCTCGGGA 265 934 CCGATGCTCTTCACCTCGGG 264 935 CCCGATGCTCTTCACCTCGG 263 936 CCCCGATGCTCTTCACCTCG 262 937 CCCCCGATGCTCTTCACCTC 261 938 GCCCCCGATGCTCTTCACCT 260 939 AGCCCCCGATGCTCTTCACC 259 940 CAGCCCCCGATGCTCTTCAC 258 941 TCAGCCCCCGATGCTCTTCA 257 942 CTCAGCCCCCGATGCTCTTC 256 943 CCTCAGCCCCCGATGCTCTT 255 944 ACCTCAGCCCCCGATGCTCT 254 945 CACCTCAGCCCCCGATGCTC 253 946 CCACCTCAGCCCCCGATGCT 252 947 CCCACCTCAGCCCCCGATGC 251 948 TCCCACCTCAGCCCCCGATG 250 949 GTCCCACCTCAGCCCCCGAT 249 950 GGTCCCACCTCAGCCCCCGA 248 951 AGGTCCCACCTCAGCCCCCG 247 952 ACAAAAACTAGCCGGGCGTGGTGGGGCACGCC 735 953 CAAAAACTAGCCGGGCGTGG 736 954 AAAAACTAGCCGGGCGTGGT 737 955 AAAACTAGCCGGGCGTGGTG 738 956 AAACTAGCCGGGCGTGGTGG 739 957 AACTAGCCGGGCGTGGTGGG 740 958 ACTAGCCGGGCGTGGTGGGG 741 959 CTAGCCGGGCGTGGTGGGGC 742 960 TAGCCGGGCGTGGTGGGGCA 743 961 AGCCGGGCGTGGTGGGGCAC 744 962 GCCGGGCGTGGTGGGGCACG 745 963 CCGGGCGTGGTGGGGCACGC 746 964 CGGGCGTGGTGGGGCACGCC 747 965 GGGCGTGGTGGGGCACGCCT 748 966 GGCGTGGTGGGGCACGCCTA 749 967 GCGTGGTGGGGCACGCCTAT 750 968 CGTGGTGGGGCACGCCTATA 751 969 GTGGTGGGGCACGCCTATAA 752 970 TGGTGGGGCACGCCTATAAT 753 971 GGTGGGGCACGCCTATAATC 754 972 GTGGGGCACGCCTATAATCC 755 973 TGGGGCACGCCTATAATCCC 756 974 GGGGCACGCCTATAATCCCA 757 975 GGGCACGCCTATAATCCCAG 758 976 GGCACGCCTATAATCCCAGC 759 977 GCACGCCTATAATCCCAGCT 760 978 CACGCCTATAATCCCAGCTT 761 979 ACGCCTATAATCCCAGCTTA 762 980 CGCCTATAATCCCAGCTTAA 763 981 TACAAAAACTAGCCGGGCGT 734 982 ATACAAAAACTAGCCGGGCG 733 983 AATACAAAAACTAGCCGGGC 732

Hot Zones (Relative upstream location to gene start site)   1-1200 1850-2200 2550-3000 3300-3500

Examples

FIG. 10 shows that BE1 (11) and BE2 (12), both at 10 μM, demonstrated statistically significant (P<0.05) inhibition compared to the untreated control inhibition values in DU145 (human prostate cell line). The negative control did not produce a statistically significant difference compared to the untreated control. The Beclin-1 sequences BE1 (11) and BE2 (12) fit the independent and dependent DNAi motif claims.

FIG. 11 shows that BE2 (12) at 10 μM demonstrated statistically significant (P<0.05) inhibition compared to the untreated and negative control values in HCT-116 (human colorectal carcinoma). The negative control did not produce a statistically significant difference compared to the untreated control. BE2 (12) fit the independent and dependent DNAi motif claims.

The secondary structures for BE1 and BE2 are shown in FIGS. 12 and 13. Sequence 11 (BE1) is shown in FIG. 12 and Sequence 12 (BE2) is shown in FIG. 13.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11951) ACTTACCACCCTCAGTGGTTTCCAGATAACATAGGCCTTCCTGAATCCCC CAGTTGAAGCAGCTCCTCCCACCCTGCCCCCACTTACTCTCTATCACATC ACCTTCTTACCTACTGTATTAGCTTTCTAGGGCTGCTGTAGCAAAGTACC ACAAAGTGGATGGCTTAGAACCAAAGAAATATATTGTCTCAGAGTTCTGG ATGCCAGAAATCCAAAATTAAGGTGTCAGCAGGACCATGTTCCTTCTAAG GGAGCCAGAGAAGTATCTGTTCCAGACCTCTTTCCTGGCTTTTGGTAGCC TCAGGTCTTCCTTGGCTTACAGATCACCCTGTGTCTCTTTACATCATCTT CCCTCAGACACGGTACATGTCTGTCTCTGTGTCCAGATTGCCCCTATTTA TAAGGACGCAGTCATATTGGTCTAGGGCTAACATCAATGACCTCATCTGC AACGATCCTATTTCCAAAAAAGGTCACATTCCCATGTGTTAGTCCCAGAT GTTAGGACTTCAACATCTTTTGGGGGACATCATTCAACCCATAATATCTG CCATTATCTGAAATTATCTTATTAACTTGGTTACATGTTTACTGTCAAAT TCTCTCCTCTGGAATATAAACTATTAGAGCAGTTCACCAGTATATCCTCT CAGACCTAGAATAGGGACTGGCACATAGTAGATGCTCAATAAACATCTGT TGAATCGATGACTGAGGATATGTTGTGTATTATTCACAATCCCTCAAGCA CTACATACACTGATTACATATACTTCCCAAGTGTGAGGATACACAGAGCA TTCACTATGTAACAGTCATTCCCCTCCATTCCAAATGTATCAGCTCATTT ATCACACTACCCTTTATGATATTTACTACTGTATACTATTAATCTCATTT TGTAAATAAGAAAACAAAGCACAGAACAGTTGAATAAATTGCATAAGGTC ACATGGTTAGTGGATGGTAAAGAACCAGGTGGTCTCAACTTCCAAATCCT CAGTTGTAACACTATACCCCCTACCTCTCTAGAAGCCCGTTACTTCTCTA TGCGTTTCTGAGATGTTAGGGACAGCCAAGCAGGAAGAAACGCAGGACTA TGAAGCAGCCACACCAGGACTAGGTGAGAATTCTTTGGGGATGATTCCAG TCACCTCCCCTAAAGGGGCTTTCATGCTGAAAGAGCCAAGAGGAAGAAGG ATTGTAAACACTATCCCTAGTCACAAAACCGGGAGAAAAATCAATCTAGT TCCACATATCACATCCAATACCAACTATAAGAAACCACATACATTTAAAA GAAAAGAAAGACACTTCTGGAGGTGGGAATAACTTTCTAAGCAGTATAAG TCATCAAGAAAAATAAGCAGATTTGACTTGAAAATTTAAAACTTCCTGAA CATCTGGAAAATAATTAAAGCATTCATGAAAAATTACTAAAAATACTGAG AAAAATACTAATAATCCAATACCTAAATAATCAAAGAATGCAAACATAAT TCAGAAAAAAGTAACTACTGCTTGAGCCCGGGAGGCGGAGATTCCAGTGA GCTGATATTGCACCACTGCACTCCAGCCTGGGTGACAGAGTGAGACCGTG TCTCTTTTTTTTTTTTTTTTTAAAAAAAGGCCGGGCATGGTGGCTCACAC CTGTAATCCCAGCACTTTGGGAGGCCAAGGCGGGCAGATCAGGAGGTCAG GAGATTGAGACCATCCTGGCTAACATGGTGAAACCGTCTCCACTAAAAAT ATAAAAAATTAGCCGGGTGTAGTGGTGGGCGCCTGTAGTCCCAGCTACTC GGGAGGCTGAGACAGGAGAATGGCGTGAACCCAGGAGGCAGAGGTTGCAG TGAGCCGAGATGGCACCACTGCACTCCAGCCTGGGCAACAGAGCAAGACT CCATCTCAAAAAAAAAAAAAAAAGTAACTACAATAAGCAAATACATAGCA AAAAGTTCAGCCTTACCAGCAATCAATGATGCTAATTAAAATAACAAGGA AGTGCCATTTTTTGCTTTTGTTCCCCAAATATATGATACCCAATACTGGC CAAGGCAATATGAAAACAGGCTTCCTCATACATTACTGGAAGCAGAATAT AGTTATGTGCAAGCACTTTGGAAAATGATTCCCAGTGTTAAGGAAGAGAC ATTAAATAGCTGACACACTCTTAATTCTGTAGTCCCAGTTATGAGTCTCT ATCATAAGTAGCCAGCTCTTCATTGCAGGATTATTGTAATCACCCACAGG GGAAATAGTAGAATTTCCAGCGGTAAAAAAATACACTAAGGCAGTACATT TAGTGTAGTGTAATGTAGCCATGATAACTACAATAACTGTGTAGCAACAT AGAAAAATGTTAAATTTAAAAAGCAGAAGCCTGGGCAACAAAGTGAGACC CCATCTCTTTTTTTTTTTGAGATGGCGTCTCGCTCTGTCACCGAGGCTGG AGTGCAGTGTGAGACCACATCTCTACAAAAAATTTTAAAAATTAGCTGGG CATGGTAGTGATCACCTGTGGTCCCTGCTACACTGGAGGTTGAAGCAAGA GGATTGCTTGAGCCAGGAAGTCAAATCTGCAGTGAGCCATGTTTGTTTGT TCCGCTTCACTCCAGCCTGGGTAACAGAGTAAGACACTGTCTCAAAATAA AAATAAAATAGACAATACTACATACAATTTTGGGTTAAGCAGTGGTTTCT TTTACACCAAAAGCATAAACATTGGACTTTATTGAAATGAAAAACTTTTG GCCAGGCACATTGGCTCACACCTGTAATCTCAGCACTTTGGGAGGCCACA GTGGGGGATTGCAAGGGGAGATGGGAAATGTTCTAAAACTGGATTATGGT GATAGTTGGGCAACTGTGTAAATTTACTAAAAATTATTGAACTGTACATT TAAAAAGTGTGAGTCTTATGGTATGTAAATTATACCCCATAAAGTTGTTT TTAAAAATGAAGTAAGTCCCTCTGCTCAAGACCCAGTCATCTCATCTCAT TCAAAGTGAAAGCCAGAGCTTTACAATCCCTATAAGAGCCTAGGTGGTAG CTCAACACTCTTACCTCCCTCACCCCATTTTCTGTATCTCTTTTCGTTGC CCATCTTCTAGCCACACCAGCCTCTGCTAATCCCCAAACAGGTACCCTCT GTGCTCTTGCTGTTCCCTTGGCCTAGAATGCTCTTCCTTAAGATGCAGGT AAGAATTCCTTCCTCACCTTCTTCAAGCTTTTATTTGAATATCACTTTCT TTTTTTGTTGGTTTTGTGTGTGTGTGTGGGGGGGGGGGGTTTGAGATGGA GTTTCCTTCTGTCGCCCAGGCTGGAGTGCAGTGGCATGATCTCGACTCAC TGCAACCTCCGCCTCCGGGGGTCAAGCGATTTTCCTACCCCAGCCTCCTG AGTAGCTGGGATTACAGGCGCACGCCACCATGCCCAGCTAATTGTATTTT TTAGTAGAGACGGGATTTAACCATTTTGGCCAGGCTGGTCTCGAACTCCT GACCTTGTGATCCGCCCGCCTCGGCCTCCCAAAGTGCTGGAATTACAAGC GTGAGCCACCATGCCCGGCCTTTTGTTGTTGCTGTTGTTGTTCTGAGATG GAGCCTTGCCCTGTCGCCCAGGCTGGAGTGCAGTGGCCCGATCTCGGCTC ACTGCAACCTCCACCTCCCAGGTTCAAGCGATTCTCCTGCCTCAGCCTCC CGAGTAGCTGGGATTAAGCTGGGATTATAGGCGTGCCCCACCACGCCCGG CTAGTTTTTGTATTTTTAGTAGAGACGGGGTTTCACTGTGTTGGCCAGGC TGGTCTCGAACTCCTGACCTCACGTGATCCGCCCTCCTCGGCCTCCCCAA GTGCTGAGATTACAGGCGTGAGCCACCGCGCCCGCCGCCCCCTGAATTTA GAGAATAGCGGAGCCTCCCCATTCTCTGCGGCCTTGGCTCCTACACTTCC CGTGGTAACCTTGTTCATCCGCTGAAGCCCGCTGCTTTTCCCAGCCCGGC CTCTGGGGGCCGCTGCCGGGCCTGTGAGCCTGTGGACCAGGAGCTCCTGC TGCCGTCGTAGCGTCACGTCCGGTCTCGGCGGAAGTTTTCCGGCGGCTAC CGGGAAGTCGCTGAAGACAGAGCGATGGTAGTTCTGGAGGCCTCGCTCCG GGGCCGACCCGAGGCCACAGTGCCTCCGCGGTAGACCGGACTTGGGTGAC GGGCTCCGGGCTCCCGAGGTGAAGAGCATCGGGGGCTGAGGTGGGACCTT AGAAGGGAGTCTGGGAACCCTCACGGCTCTTATTGGAGTCCCTTCCCTGA CCCTGGGCTCTAAACTGCCTTTGCTCAGGCTGTCCCGGAAGCAGGTCCTC CCCGTATCATACCATTCCTAGAGGGAACGGCGCAGGTTGGGACTTCCCTC CCTTTACCATCGTCACCAAGGCATGTGGTAACCCCGGGCCGGAGGTCAAG GGCGTCGCTTCTCCCTAATGTTGCCTCTTTTCCACGGCCTCAGGG ATG

3) STAT3. Signal Transducers and Activators of Transcription 3 (STAT3) is a point of convergence for numerous oncogenic signalling pathways, is constitutively activated both in tumor cells and in immune cells in the tumor microenvironment. STAT3 inhibits the expression of mediators necessary for immune activation against tumor cells (Nature Reviews Immunology 7, 41-51; 2007; Proc Natl Acad Sci USA. 2006 Jul. 5; 103(27): 10151-10152) and promotes the production of immunosuppressive factors that further activate STAT3 in diverse immune-cell subsets, altering gene-expression. This restraining anti-tumor immune response and propagation of cross-talk between tumor cells and their immunological microenvironment leads to tumor-induced immunosuppression and enhanced tumor growth. STAT3 belongs to a protein family of transcription factors first characterized for their role in cytokine signaling that contain a site for specific tyrosine phosphorylation, a modification that results in a conformational rearrangement causing it to accumulate in the cell nucleus, bound to enhancer elements of target genes (Nat. Rev. Mol. Cell. Biol. 2002; 3:651-662). STAT3 is a substrate for the catalytic activity of the tyrosine kinase oncoprotein v-Src (Science. 1995; 269:81-83) and that phosphorylated STAT3 accumulated in many human cancers, suggesting that activated STAT3 may act as an oncogene (Cell. 1999; 98:295-303). In a recent issue of PNAS, Kasprzycka et al. (Proc. Natl. Acad. Sci. USA. 2006; 103:9964-9969) provided evidence that activated STAT3 in a tumor cell contributes to both cell survival and impaired immune surveillance by conferring properties of a T lymphocyte regulatory phenotype on a T cell lymphoma. Further it is recognized that STAT3 is stimulated by classic growth-promoting signals, such as activated growth factor receptors as well as a remarkable degree of diversity for the molecular mechanisms at the basis of STAT3 action including some noncanonical mechanisms of tumor progression that apparently do not rely on tyrosine phosphorylation or binding of homodimers to DNA (Cancer Res. 2005; 65:939-947), possibly involving pathways in malignant cells not directly regulating gene expression.

Isis Pharmaceuticals is developing an antisense against STAT3. In preclinical studies, ISIS-STAT3Rx demonstrated antitumor activity in animal models of human cancer. ISIS-STAT3Rx was tested in a Phase 1 study in patients with solid tumors and lymphoma who have relapsed or were refractory to multiple chemotherapy regimens and in a Phase 2 study in focused patient populations with advanced cancers that have been linked to STAT3 and who have failed all other treatment options with clear responses in patients with advanced cancer who were refractory to prior chemotherapy treatment. STAT3 is implicated in a variety of cancers, including brain, lung, breast, bone, liver and multiple myeloma to promote tumor cell growth and prevents cell death.

Protein: STAT3 Gene: STAT3 (Homo sapiens, chromosome 17, 40465343-40540513 [NCBI Reference Sequence: NC_(—)000017.10]; start site location: 40540405; strand: negative)

Gene Identification GeneID 6774 HGNC 11364 HPRD 00026 MIM 102582

Targeted Sequences Relative upstream location to gene start site Sequence Design (upstream promoter of the ID No: ID Sequence (5′-3′) two promoters) 984 ST1 GGCCGAGGCACGCCGTCATGCA −18 985 ST2 CCGGCCCTTGGCACCACGTGGTGGCGA 345 986 TTGTTCCCTCGGCTGCGACGTCG −135 987 CAGTCTGCGCCGCCGCAGCTCCGG −92 988 CAGTGCGTGTGCGGTACAGCCG 45 989 TGTGCTGGCTGTTCCGACAGTTCGGT 140 990 TAACTACGCTATCCCGTGCGGCC 1998449 991 TCGCCCAGCCCCAGCCTGGCCGAGGC −35

Hot Zones (Relative upstream location to gene start site) −200-200  300-400 1998400-1998500

Examples

FIG. 14 shows ST1 (21) and ST2 (22), both at 10 μM, demonstrated statistically significant (P<0.05) inhibition compared to the untreated control inhibition values in MDA-MB-231 (human breast cell line). The negative control did not produce a statistically significant difference compared to the untreated control. The STAT3 sequence ST2 (22) fit the independent and dependent DNAi motif claims. The STAT3 sequence ST1 (21) is designed to the coding region of STAT3.

FIG. 15, which is similar to FIG. 12, shows ST1 (21) and ST2 (22), both at 10 μM, demonstrated statistically significant (P<0.05) inhibition compared to the untreated control inhibition values in DU145 (human prostate cell line). The negative control did not produce statistically significant difference compared to the untreated control. The STAT3 sequence ST2 (22) fit the independent and dependent DNAi motif claims. The STAT3 sequence ST1 (21) is designed to the coding region of STAT3.

The secondary structures for ST1 and ST2 are shown in FIGS. 16 and 17. Sequence 21 (ST1) is shown in FIG. 16 and Sequence 22 (ST2) is shown in FIG. 17.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11952) CTTCTGCACTTAAGCACACTATACTTTTTTCACCCAAAGTACCAAATCAA ACTAGTCAGGATACCTACCTTTGTACAATGTCAGACTCCAGTTAATAACT CCCCTAGGGCAGAGGGCATATGCACTGATTTACTTTGTACAAATTAACCA GCATCAGGCAATCAGGCCTGTGCCTAACACATAGTAAGCACTCTATGATT AAACATCAGTGCTTCGGCTCCAAAGTTTTATTTATTTATTTATTTATTTA TTTTTTTTTTTTTTGAGACGGAGTCTCGCTCTGTCGCCCAGGCTGGAGTG CAGTGGTGCGATATCGGCTCACTGCAAGCTCCGCCTCCCGAGTTCACGCT CTTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGACGCCCGCCAC AACGCCCGGCTACTTTTTTTTGTATTTTTAGTAGAGATGGGGTTTCACCG TGTTAGCCAGGATGGTCTCGATCTCCTGGCCTCGTGATCCGCGCGTCTGG GCCTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCGCGCCCGGCGCCC CGAAAGTTTTAAAAGCTTCCCCTACAAAAGAACAGAACTGAAATTCCTTG GTCCTGTATTCAATGTCTTTTGTAAGTAATCACTTCTCCCCTACTTACCC TCCTAGTCTACCGGGCTACCAGGAATTTTTTTTTTTTTTGGAGACAGGGT CTCACTCTGTCACCCAGGCTGGAGTGCGGTGGCGGGATCACGGCTCACTG CAGCCTTAACCCCCGGGGCTTGGGTGATCCTCCCACCTTAGTCTCACCAG TAGCTGGGACTACAGGTCCACGCCACCAGGCCTGGCTAATTTTTTTTATT TTTAGGGGAGAGGGAGTTTTACCACGTTGCCCAAGCTGGTCTCAAACTCC TGGGCTCAAGCAATCCTCCTGCCTCAGCCTCCCAAAGTGCTGGGATTACA GGCATAAACCACCGCAAATTCTTTACACCTATCAAATTCCACCCATTATT TGGGACCCAGTTGAAATCCCTCTTTGGCAAAAAGACTTTCTAGACAACTC CAGGCCTCATAACCTCTCCTTTCTCTGAAGATCTGTAGCATTCAGCCTAG CACTGTCCAATAGAACGTTCTATGATAACAGAAAAGTTCTACATCTGTAC TGTATGTTCTTTTATGTAGAACAGCTACCTTGTTAGCACAAGTGTAAAGT CTCACCATCTCTTTGATGACAACATGTTACATTGGATGGTTAAAACATTT ATCAGCTCCCCCAGTAGACTGCAATTTCTGTGAACAAGATACAACTTATT CTTCATAGCAACTCTGACAAAGTTGCAAAAGGTATATATATGTTGGCCAG GCAAGGTGGTTCACGCCTGTAATCCCAGCACTTTGGGAGGCTGAGGTGGG CAGATCTCTTGAGGTCAGGAGTTTGAGACCAGCCTGGTCAATATAGTGAA ACCTTATCTCTACTAAAAATACAAAAATTAGCCGGGCGTAGTGGCGGGCA CCTGTAATCCCAGCTACTCAGGAGGCTGAGGTGCGAGAATCACTTGAACC CGGGAGGAGGAGGTTGCAGTGAGCCACGATCATGCCACTGCACTCCAGCC TGGGTGATAGAGTGCAACTCCAACTCAAAAAAAAAAAAAAAAAAGTATAT ATTTGTTGATTTGCACATCACCTAAGAAAACCATAAGCTAAGAAGGTTTG GACTCAGGCGTCTGGAAAGTTGGTCACCACCTCTACCCCACCTCATATCT GAATGTCAAGAGACACGTAGAGGCAGAGAAGTTAAAGCAACTTTCTAGAG ACAGAAATGACCACTGATCAAGCCACAATGCACTCTGGTTTAAATGACAT TTAGGTCATGACTGTCCTTAATCTAAAACAAACCTAGATTAGTATTTCTT TTCATTAGTAAATAGCTAAATTCTGATGGTAAATTATGCTGACCAAAAAC AGTTCCTCACTTCCCAAGTTAGACATAGCAATTAGAAAAATAATCTAAGC AAGCTCCATTTGTATTTCTTTTTTCACCTGTTTATTGAATATTTACCTCC CATGAAGTCTTTCAGCCTATTGGTGGTATTTTACTGTTCAGATATATGTT AGAATTTCACTGATACTTACTGGGCGCGGTGGCTCACACCTGTAATCCCA GCACTTTGGGAGATAGAGGTGGGCAAATCACAAGGTCAGGAGTTCAAGAC CAGCCTGGCCAATATGGTAAAACCCCGGTCTCTACTAAAAATACAAAAAT TAGCTGGGCGTGGTGGCGCACGCCTGTAGTCCCAGCTACTTGGGAGGCTG AGGCAGGAGAATCGCTTGAACCCAGGAGGCAGAGGTTGCAGTGAGCCAAG ATTGCGCCACTGCACTCTAGCCTGGGCAACAGAGCAAGACTCTGACTCAA AAAAAAAAAAAAAAAGAATTTCACTGATACTTTTCACAAAATATACAGAA GGAGGCACAAATTCCACCACTATGGCACTCTGCTGCGTTGGCCAAGTGTC TTGATCCTTTGGCCTCAATTTTCTTATCTACGATATTAGGGTAATTGTTA TGTGAACTACCCACCTCACAAGTCCTTTGTGGGTTAATTCATAACTGTGC TGTGGGTATTTCTTTTTCTTTCCTTTCTTCCTCCTTTCCTTTCTTTCTTT CTTAAAGATGGGTTCTCATTATGGTGCTTAGACTAGACTCTAGACCCAAT TCCTGGCCTCTCACCATGTTGCCCAGACCAGACTCAACTCCTGGACTCAA GGAATCCTCCCACCTCAGCCTTCAATTAGCTGGGATCAGAGGTGTGCACC ACCATGCCTGGCACTGTGGATATTTCTAAGTGATTATTCTTCTCAAATGA ACTACATAAAAAACAAAAGATTCATGAATTTACTAATGGTTCTTTGTGAT GGATGTGCTAATATAGAGACTAAAATCAAGGCTCCAACCTCTAAAACATT TTTTTTTAAATTCCAGACTTGTTTCCCCATCCCACTGTGCAAACTGAACA AAAACTGGGCTAGCACTCCTGTCTGGAACATGTAATAAGGAAATAAATGT GCTGACTCAGAGAACACAGACATATTTAATATAAAATAAGATAGAAAACT GGCTGAACCAAGTCATAACACAGTCTAAATCCACATATAAAAGATTGAGA TGATTTTCTGCTTTGCTTTATTCAAGCCCAATGCTTTATCAGCACAGCCA GCCAAAAATTTACAACCCATACACAGACTATGTAAACCTTTAGTTGCACA TACAGTAAGACCAGCAGGTACACACTATACACATTTTTAATTAAAAAAAT GACTAACCACTGATTTTGTCACCACACTTAACAACGACCTGATATGGCAC AGAGTGATGTGTACCAAACATGGAAATACCAACTTGGGCGACGGTTTGAA TCTTGTTAACTTCAGTGCAACCACACCCCCCAAATGCATGTAAAGTTTGC ACACATGGTTTTTTCAAGGCCAGCCTGTCTTTGTTTCCCTCTCCTCTGCA TTTACCCAAGATCTTGGCTCTGAGACAGAAAACTCCCACTCTCAATTGGT TCATTCCGTCCTATGCAATTAAGCAACACCACAATCCAGTAAATGCAATG GCTCAATTATTTATCTTCTGGCCGACTTTACCAGGTATTTGGAAAAGGAC AATGTCAAGAGGTTTATTTCTCTCTCTAGAGCTGGCTTGACGGGTTGATG GGGATTTTATTTTGTCTTTTTTTCTCTTTTTTACAAGGCGGGGACGTGGG GGGAGCATAATTTAACCTAGAAAAAGATGCGAGGGAATTTAGAAAGAGTA CCGGTCTGTCAATTTCCCTACAGGAAACTTGATTCTTATGCAATAAAGCC TACCCACGACCAGCCAGCCCGTAAGGCTGCAGGCGACAGACACACCTATT CCTGCCTCCAAAAGGGCACAGCTGTCTCCTGAAGGAGCGGGAACAGGGCA AGCGGAGGAAGTGGCTCAGCGGGAGCCGCCGACCGGGCGGGGAGGAGGCG CTTTCCGACCCCCCACTCGCGCCGGTGATCCCCGTCGGCGTGACAGTCGC TCCGGTGGCCGGAACGTCCCCAGGGCCCCAGGGAGCAGGAAATCGGGGGA CTGTCCCTCACTCCTGCCGCCGCAACCGAGTGCGCCCTCGCCCCACGGTG CCCCCTCGAGCGCGTTCTGTTTCTCCGAAGAACGAAACTTCCCTCCAGCG CCCCGAGTCCCTTCCGAGGCCCGCTCCTGTCATCCCGAAGAGTCTTCCCT CAGGGCGACCCTCCGCGTCTCTTCATCTCTCCCGGCCCCACTGCAGCGTC CATCACAACATCCCCAAGGTCCCAGAGGCCCCCTGCCGCTGCGGAGCCCC CGGGTCCCCAGGCCTCCCCAACGGCCCCACCCTGCACCCCCTTCACCTGT TTCTCCGGCAGAGGCCGAGAGGCCGGGGCTGCGCGTGTGCCGGGGACGGG CGGCGAGGCTCCCTCAGGCCGAAGGGCCTCTCCGAGCCGAGGGGGAGAGA CAGCGCC

4) HIF1A. Hypoxia-inducible factors (HIFs) are transcription factors that respond to changes in available oxygen in the cellular environment, specifically, to decreases in oxygen, or hypoxia. Hypoxia-inducible factor-1 (HIF-1a) is the alpha subunit of the HIF-1 dimeric transcriptional complex involved in the maintenance of oxygen and energy homoeostasis. Hypoxia often keeps cells from differentiating. However, hypoxia promotes the formation of blood vessels, and is important for the formation of a vascular system in embryos, and cancer tumors. The HIF-1 alpha subunit is oxygen labile and is degraded by the proteasome following prolyl-hydroxylation and ubiquitination in normoxic cells. There is also evidence that HIF-1 is also involved in immune reactions (Hurwig-Burgel et al, J Interferon Cytokine Res. 2005; 25(6):297-310). Immunomodulatory peptides, including interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha), stimulate HIF-1 dependent gene expression even in normoxic cells. Both the hypoxic and the cytokine-induced activation of HIF-1 involve the phosphatidylinositol-3-kinase (PI3K) and the mitogen-activated protein kinase (MAPK) signaling pathways. In addition, heat shock proteins (HSP) and other cofactors interact with HIF-1 subunits. HIF-1 blockade may be beneficial to prevent tumor angiogenesis and tumor growth.

Protein: HIF1A Gene: HIF1A (Homo sapiens, chromosome 14, 62162119-62214977 [NCBI Reference Sequence: NC_(—)000014.8]; start site location: 62162523; strand: positive)

Gene Identification GeneID 3091 HGNC 4910 HPRD 04517 MIM 603348

Targeted Sequences Relative upstream Sequence location to gene start ID No: Design ID Sequence (5′-3′) site 992 HI1 CAGGCCGGCGCGCGCTCCCGCAA 390 1048 HI2 GGACGGGCTGCGACGCTCACGTGC 539 1089 GAGGTGGGGGTGCGAGGCGGGAAACCC 108 CTCG 1090 CAATCGCCGGGGTCCGGGCCCGGC 162 1129 TGGCCGAAGCGACGAAGAGGG 232 1130 GGGCGGAGGCGCGCTCGGGCGCG 325 1142 CACGGCGGGCGGCCCCCAGGCTCGC 26 1214 CAGGCCGGCGCGCGCTCCCGCAAGCCCG 390 1270 CGATTGCCGCCCAACTCTGCTGGG 789

Target Shift Sequences Relative upstream location to Sequence gene start ID No: Sequence (5′-3′) site 992 CAGGCCGGCGCGCGCTCCCGCAA 390 993 AGGCCGGCGCGCGCTCCCGC 391 994 GGCCGGCGCGCGCTCCCGCA 392 995 GCCGGCGCGCGCTCCCGCAA 393 996 CCGGCGCGCGCTCCCGCAAG 394 997 CGGCGCGCGCTCCCGCAAGC 395 998 GGCGCGCGCTCCCGCAAGCC 396 999 GCGCGCGCTCCCGCAAGCCC 397 1000 CGCGCGCTCCCGCAAGCCCG 398 1001 GCGCGCTCCCGCAAGCCCGC 399 1002 CGCGCTCCCGCAAGCCCGCC 400 1003 GCGCTCCCGCAAGCCCGCCT 401 1004 CGCTCCCGCAAGCCCGCCTC 402 1005 GCTCCCGCAAGCCCGCCTCA 403 1006 CTCCCGCAAGCCCGCCTCAC 404 1007 TCCCGCAAGCCCGCCTCACC 405 1008 CCCGCAAGCCCGCCTCACCT 406 1009 CCGCAAGCCCGCCTCACCTG 407 1010 CGCAAGCCCGCCTCACCTGA 408 1011 GCAAGCCCGCCTCACCTGAG 409 1012 CAAGCCCGCCTCACCTGAGG 410 1013 AAGCCCGCCTCACCTGAGGT 411 1014 AGCCCGCCTCACCTGAGGTG 412 1015 GCCCGCCTCACCTGAGGTGG 413 1016 CCCGCCTCACCTGAGGTGGA 414 1017 CCGCCTCACCTGAGGTGGAG 415 1018 CGCCTCACCTGAGGTGGAGG 416 1019 CCAGGCCGGCGCGCGCTCCC 389 1020 CCCAGGCCGGCGCGCGCTCC 388 1021 GCCCAGGCCGGCGCGCGCTC 387 1022 TGCCCAGGCCGGCGCGCGCT 386 1023 CTGCCCAGGCCGGCGCGCGC 385 1024 CCTGCCCAGGCCGGCGCGCG 384 1025 GCCTGCCCAGGCCGGCGCGC 383 1026 CGCCTGCCCAGGCCGGCGCG 382 1027 TCGCCTGCCCAGGCCGGCGC 381 1028 CTCGCCTGCCCAGGCCGGCG 380 1029 GCTCGCCTGCCCAGGCCGGC 379 1030 CGCTCGCCTGCCCAGGCCGG 378 1031 CCGCTCGCCTGCCCAGGCCG 377 1032 CCCGCTCGCCTGCCCAGGCC 376 1033 GCCCGCTCGCCTGCCCAGGC 375 1034 CGCCCGCTCGCCTGCCCAGG 374 1035 GCGCCCGCTCGCCTGCCCAG 373 1036 CGCGCCCGCTCGCCTGCCCA 372 1037 GCGCGCCCGCTCGCCTGCCC 371 1038 AGCGCGCCCGCTCGCCTGCC 370 1039 GAGCGCGCCCGCTCGCCTGC 369 1040 GGAGCGCGCCCGCTCGCCTG 368 1041 GGGAGCGCGCCCGCTCGCCT 367 1042 CGGGAGCGCGCCCGCTCGCC 366 1043 GCGGGAGCGCGCCCGCTCGC 365 1044 GGCGGGAGCGCGCCCGCTCG 364 1045 GGGCGGGAGCGCGCCCGCTC 363 1046 GGGGCGGGAGCGCGCCCGCT 362 1047 GGGGGCGGGAGCGCGCCCGC 361 1048 GGACGGGCTGCGACGCTCACGTGC 539 1049 GACGGGCTGCGACGCTCACG 540 1050 ACGGGCTGCGACGCTCACGT 541 1051 CGGGCTGCGACGCTCACGTG 542 1052 GGGCTGCGACGCTCACGTGC 543 1053 GGCTGCGACGCTCACGTGCT 544 1054 GCTGCGACGCTCACGTGCTC 545 1055 CTGCGACGCTCACGTGCTCG 546 1056 TGCGACGCTCACGTGCTCGT 547 1057 GCGACGCTCACGTGCTCGTC 548 1058 CGACGCTCACGTGCTCGTCT 549 1059 GACGCTCACGTGCTCGTCTG 550 1060 ACGCTCACGTGCTCGTCTGT 551 1061 CGCTCACGTGCTCGTCTGTG 552 1062 GCTCACGTGCTCGTCTGTGT 553 1063 CTCACGTGCTCGTCTGTGTT 554 1064 TCACGTGCTCGTCTGTGTTT 555 1065 CACGTGCTCGTCTGTGTTTA 556 1066 ACGTGCTCGTCTGTGTTTAG 557 1067 CGTGCTCGTCTGTGTTTAGC 558 1068 GTGCTCGTCTGTGTTTAGCG 559 1069 TGCTCGTCTGTGTTTAGCGG 560 1070 GCTCGTCTGTGTTTAGCGGC 561 1071 CTCGTCTGTGTTTAGCGGCG 562 1072 TCGTCTGTGTTTAGCGGCGG 563 1073 CGTCTGTGTTTAGCGGCGGA 564 1074 GTCTGTGTTTAGCGGCGGAG 565 1075 TCTGTGTTTAGCGGCGGAGG 566 1076 CTGTGTTTAGCGGCGGAGGA 567 1077 TGTGTTTAGCGGCGGAGGAA 568 1078 GGGACGGGCTGCGACGCTCA 538 1079 TGGGACGGGCTGCGACGCTC 537 1080 CTGGGACGGGCTGCGACGCT 536 1081 GCTGGGACGGGCTGCGACGC 535 1082 AGCTGGGACGGGCTGCGACG 534 1083 CAGCTGGGACGGGCTGCGAC 533 1084 ACAGCTGGGACGGGCTGCGA 532 1085 CACAGCTGGGACGGGCTGCG 531 1086 GCACAGCTGGGACGGGCTGC 530 1087 GGCACAGCTGGGACGGGCTG 529 1088 AGGCACAGCTGGGACGGGCT 528 1089 GAGGTGGGGGTGCGAGGCGGGAAACCCCTCG 108 1090 CAATCGCCGGGGTCCGGGCCCGGC 162 1091 AATCGCCGGGGTCCGGGCCC 163 1092 ATCGCCGGGGTCCGGGCCCG 164 1093 TCGCCGGGGTCCGGGCCCGG 165 1094 CGCCGGGGTCCGGGCCCGGC 166 1095 GCCGGGGTCCGGGCCCGGCT 167 1096 CCGGGGTCCGGGCCCGGCTC 168 1097 CGGGGTCCGGGCCCGGCTCC 169 1098 GGGGTCCGGGCCCGGCTCCG 170 1099 GGGTCCGGGCCCGGCTCCGA 171 1100 GGTCCGGGCCCGGCTCCGAG 172 1101 GTCCGGGCCCGGCTCCGAGC 173 1102 TCCGGGCCCGGCTCCGAGCC 174 1103 CCGGGCCCGGCTCCGAGCCT 175 1104 CGGGCCCGGCTCCGAGCCTC 176 1105 GGGCCCGGCTCCGAGCCTCT 177 1106 GGCCCGGCTCCGAGCCTCTC 178 1107 GCCCGGCTCCGAGCCTCTCC 179 1108 CCCGGCTCCGAGCCTCTCCT 180 1109 CCGGCTCCGAGCCTCTCCTC 181 1110 CGGCTCCGAGCCTCTCCTCA 182 1111 GGCTCCGAGCCTCTCCTCAG 183 1112 GCTCCGAGCCTCTCCTCAGG 184 1113 CTCCGAGCCTCTCCTCAGGT 185 1114 TCCGAGCCTCTCCTCAGGTG 186 1115 CCGAGCCTCTCCTCAGGTGG 187 1116 CGAGCCTCTCCTCAGGTGGC 188 1117 GCAATCGCCGGGGTCCGGGC 161 1118 GGCAATCGCCGGGGTCCGGG 160 1119 CGGCAATCGCCGGGGTCCGG 159 1120 GCGGCAATCGCCGGGGTCCG 158 1121 GGCGGCAATCGCCGGGGTCC 157 1122 GGGCGGCAATCGCCGGGGTC 156 1123 CGGGCGGCAATCGCCGGGGT 155 1124 GCGGGCGGCAATCGCCGGGG 154 1125 AGCGGGCGGCAATCGCCGGG 153 1126 AAGCGGGCGGCAATCGCCGG 152 1127 GAAGCGGGCGGCAATCGCCG 151 1128 AGAAGCGGGCGGCAATCGCC 150 1129 TGGCCGAAGCGACGAAGAGGG 232 1130 GGGCGGAGGCGCGCTCGGGCGCG 325 1131 GGCGGAGGCGCGCTCGGGCG 326 1132 GCGGAGGCGCGCTCGGGCGC 327 1133 CGGAGGCGCGCTCGGGCGCG 328 1134 GGAGGCGCGCTCGGGCGCGC 329 1135 GAGGCGCGCTCGGGCGCGCG 330 1136 AGGCGCGCTCGGGCGCGCGG 331 1137 GGCGCGCTCGGGCGCGCGGG 332 1138 GCGCGCTCGGGCGCGCGGGG 333 1139 CGCGCTCGGGCGCGCGGGGA 334 1140 GCGCTCGGGCGCGCGGGGAG 335 1141 CGCTCGGGCGCGCGGGGAGG 336 1142 CACGGCGGGCGGCCCCCAGGCTCGC 26 1143 ACGGCGGGCGGCCCCCAGGC 27 1144 CGGCGGGCGGCCCCCAGGCT 28 1145 GGCGGGCGGCCCCCAGGCTC 29 1146 GCGGGCGGCCCCCAGGCTCG 30 1147 CGGGCGGCCCCCAGGCTCGC 31 1148 GGGCGGCCCCCAGGCTCGCT 32 1149 GGCGGCCCCCAGGCTCGCTC 33 1150 GCGGCCCCCAGGCTCGCTCC 34 1151 CGGCCCCCAGGCTCGCTCCG 35 1152 GGCCCCCAGGCTCGCTCCGG 36 1153 GCCCCCAGGCTCGCTCCGGC 37 1154 CCCCCAGGCTCGCTCCGGCC 38 1155 CCCCAGGCTCGCTCCGGCCT 39 1156 CCCAGGCTCGCTCCGGCCTA 40 1157 CCAGGCTCGCTCCGGCCTAA 41 1158 CAGGCTCGCTCCGGCCTAAG 42 1159 AGGCTCGCTCCGGCCTAAGC 43 1160 GGCTCGCTCCGGCCTAAGCG 44 1161 GCTCGCTCCGGCCTAAGCGC 45 1162 CTCGCTCCGGCCTAAGCGCT 46 1163 TCGCTCCGGCCTAAGCGCTG 47 1164 CGCTCCGGCCTAAGCGCTGG 48 1165 GCTCCGGCCTAAGCGCTGGC 49 1166 CTCCGGCCTAAGCGCTGGCT 50 1167 TCCGGCCTAAGCGCTGGCTC 51 1168 CCGGCCTAAGCGCTGGCTCC 52 1169 CGGCCTAAGCGCTGGCTCCC 53 1170 GGCCTAAGCGCTGGCTCCCT 54 1171 GCCTAAGCGCTGGCTCCCTC 55 1172 CCTAAGCGCTGGCTCCCTCC 56 1173 CTAAGCGCTGGCTCCCTCCA 57 1174 TAAGCGCTGGCTCCCTCCAC 58 1175 AAGCGCTGGCTCCCTCCACA 59 1176 AGCGCTGGCTCCCTCCACAC 60 1177 GCGCTGGCTCCCTCCACACG 61 1178 CGCTGGCTCCCTCCACACGC 62 1179 GCTGGCTCCCTCCACACGCG 63 1180 CTGGCTCCCTCCACACGCGG 64 1181 TGGCTCCCTCCACACGCGGA 65 1182 GGCTCCCTCCACACGCGGAG 66 1183 GCTCCCTCCACACGCGGAGA 67 1184 CTCCCTCCACACGCGGAGAA 68 1185 TCCCTCCACACGCGGAGAAG 69 1186 CCCTCCACACGCGGAGAAGA 70 1187 CCTCCACACGCGGAGAAGAG 71 1188 CTCCACACGCGGAGAAGAGA 72 1189 TCACGGCGGGCGGCCCCCAG 25 1190 TTCACGGCGGGCGGCCCCCA 24 1191 CTTCACGGCGGGCGGCCCCC 23 1192 TCTTCACGGCGGGCGGCCCC 22 1193 GTCTTCACGGCGGGCGGCCC 21 1194 TGTCTTCACGGCGGGCGGCC 20 1195 ATGTCTTCACGGCGGGCGGC 19 1196 GATGTCTTCACGGCGGGCGG 18 1197 CGATGTCTTCACGGCGGGCG 17 1198 GCGATGTCTTCACGGCGGGC 16 1199 CGCGATGTCTTCACGGCGGG 15 1200 CCGCGATGTCTTCACGGCGG 14 1201 CCCGCGATGTCTTCACGGCG 13 1202 CCCCGCGATGTCTTCACGGC 12 1203 TCCCCGCGATGTCTTCACGG 11 1204 GTCCCCGCGATGTCTTCACG 10 1205 GGTCCCCGCGATGTCTTCAC 9 1206 CGGTCCCCGCGATGTCTTCA 8 1207 TCGGTCCCCGCGATGTCTTC 7 1208 ATCGGTCCCCGCGATGTCTT 6 1209 AATCGGTCCCCGCGATGTCT 5 1210 GAATCGGTCCCCGCGATGTC 4 1211 TGAATCGGTCCCCGCGATGT 3 1212 GTGAATCGGTCCCCGCGATG 2 1213 GGTGAATCGGTCCCCGCGAT 1 1214 CAGGCCGGCGCGCGCTCCCGCAAGCCCG 390 1215 AGGCCGGCGCGCGCTCCCGC 391 1216 GGCCGGCGCGCGCTCCCGCA 392 1217 GCCGGCGCGCGCTCCCGCAA 393 1218 CCGGCGCGCGCTCCCGCAAG 394 1219 CGGCGCGCGCTCCCGCAAGC 395 1220 GGCGCGCGCTCCCGCAAGCC 396 1221 GCGCGCGCTCCCGCAAGCCC 397 1222 CGCGCGCTCCCGCAAGCCCG 398 1223 GCGCGCTCCCGCAAGCCCGC 399 1224 CGCGCTCCCGCAAGCCCGCC 400 1225 GCGCTCCCGCAAGCCCGCCT 401 1226 CGCTCCCGCAAGCCCGCCTC 402 1227 GCTCCCGCAAGCCCGCCTCA 403 1228 CTCCCGCAAGCCCGCCTCAC 404 1229 TCCCGCAAGCCCGCCTCACC 405 1230 CCCGCAAGCCCGCCTCACCT 406 1231 CCGCAAGCCCGCCTCACCTG 407 1232 CGCAAGCCCGCCTCACCTGA 408 1233 GCAAGCCCGCCTCACCTGAG 409 1234 CAAGCCCGCCTCACCTGAGG 410 1235 AAGCCCGCCTCACCTGAGGT 411 1236 AGCCCGCCTCACCTGAGGTG 412 1237 GCCCGCCTCACCTGAGGTGG 413 1238 CCCGCCTCACCTGAGGTGGA 414 1239 CCGCCTCACCTGAGGTGGAG 415 1240 CGCCTCACCTGAGGTGGAGG 416 1241 CCAGGCCGGCGCGCGCTCCC 389 1242 CCCAGGCCGGCGCGCGCTCC 388 1243 GCCCAGGCCGGCGCGCGCTC 387 1244 TGCCCAGGCCGGCGCGCGCT 386 1245 CTGCCCAGGCCGGCGCGCGC 385 1246 CCTGCCCAGGCCGGCGCGCG 384 1247 GCCTGCCCAGGCCGGCGCGC 383 1248 CGCCTGCCCAGGCCGGCGCG 382 1249 TCGCCTGCCCAGGCCGGCGC 381 1250 CTCGCCTGCCCAGGCCGGCG 380 1251 GCTCGCCTGCCCAGGCCGGC 379 1252 CGCTCGCCTGCCCAGGCCGG 378 1253 CCGCTCGCCTGCCCAGGCCG 377 1254 CCCGCTCGCCTGCCCAGGCC 376 1255 GCCCGCTCGCCTGCCCAGGC 375 1256 CGCCCGCTCGCCTGCCCAGG 374 1257 GCGCCCGCTCGCCTGCCCAG 373 1258 CGCGCCCGCTCGCCTGCCCA 372 1259 GCGCGCCCGCTCGCCTGCCC 371 1260 AGCGCGCCCGCTCGCCTGCC 370 1261 GAGCGCGCCCGCTCGCCTGC 369 1262 GGAGCGCGCCCGCTCGCCTG 368 1263 GGGAGCGCGCCCGCTCGCCT 367 1264 CGGGAGCGCGCCCGCTCGCC 366 1265 GCGGGAGCGCGCCCGCTCGC 365 1266 GGCGGGAGCGCGCCCGCTCG 364 1267 GGGCGGGAGCGCGCCCGCTC 363 1268 GGGGCGGGAGCGCGCCCGCT 362 1269 GGGGGCGGGAGCGCGCCCGC 361 1270 CGATTGCCGCCCAACTCTGCTGGG 789 1271 GATTGCCGCCCAACTCTGCT 790 1272 ATTGCCGCCCAACTCTGCTG 791 1273 TTGCCGCCCAACTCTGCTGG 792 1274 TGCCGCCCAACTCTGCTGGG 793 1275 GCCGCCCAACTCTGCTGGGC 794 1276 CCGCCCAACTCTGCTGGGCT 795 1277 CGCCCAACTCTGCTGGGCTC 796 1278 ACGATTGCCGCCCAACTCTG 788 1279 CACGATTGCCGCCCAACTCT 787 1280 GCACGATTGCCGCCCAACTC 786 1281 GGCACGATTGCCGCCCAACT 785 1282 GGGCACGATTGCCGCCCAAC 784 1283 TGGGCACGATTGCCGCCCAA 783 1284 CTGGGCACGATTGCCGCCCA 782 1285 GCTGGGCACGATTGCCGCCC 781 1286 TGCTGGGCACGATTGCCGCC 780 1287 GTGCTGGGCACGATTGCCGC 779 1288 AGTGCTGGGCACGATTGCCG 778 1289 CAGTGCTGGGCACGATTGCC 777 1290 TCAGTGCTGGGCACGATTGC 776 1291 CTCAGTGCTGGGCACGATTG 775 1292 CCTCAGTGCTGGGCACGATT 774 1293 GCCTCAGTGCTGGGCACGAT 773 1294 GGCCTCAGTGCTGGGCACGA 772 1295 CGGCCTCAGTGCTGGGCACG 771 1296 TCGGCCTCAGTGCTGGGCAC 770 1297 CTCGGCCTCAGTGCTGGGCA 769 1298 CCTCGGCCTCAGTGCTGGGC 768 1299 TCCTCGGCCTCAGTGCTGGG 767 1300 CTCCTCGGCCTCAGTGCTGG 766 1301 TCTCCTCGGCCTCAGTGCTG 765 1302 TTCTCCTCGGCCTCAGTGCT 764 1303 TTTCTCCTCGGCCTCAGTGC 763 1304 CTTTCTCCTCGGCCTCAGTG 762 1305 TCTTTCTCCTCGGCCTCAGT 761 1306 CTCTTTCTCCTCGGCCTCAG 760 1307 TCTCTTTCTCCTCGGCCTCA 759 1308 CTCTCTTTCTCCTCGGCCTC 758 1309 GCTCTCTTTCTCCTCGGCCT 757 1310 TGCTCTCTTTCTCCTCGGCC 756 1311 CTGCTCTCTTTCTCCTCGGC 755 1312 CCTGCTCTCTTTCTCCTCGG 754 1313 TCCTGCTCTCTTTCTCCTCG 753

Hot Zones (Relative upstream location to gene start site)   1-1050 1500-1700 2000-2450

FIG. 18 shows MDA-MB-231 (human breast cell line), HI1 (31) and HI2 (32) at 10 μM showed increased inhibition compared to the untreated control and the negative control. The HIF1A sequences HI1 (31) and HI2 (32) (shown below) fit the independent and dependent DNAi motif claims.

FIG. 19 shows DU145 (human prostate cell line), HI1 (31) and HI2 (32) at 10 μM produced statistically significant (P<0.05) inhibition compared to the untreated control values. The negative control inhibition values did not a produce statistically significant difference compared to the untreated control values. The HIF1A sequences HI1 (31) and HI2 (32) (shown below) fit the independent and dependent DNAi motif claims.

The secondary structures for HI1 and HI2 are shown in FIGS. 20 and 21. Sequence 31 (HI1) is shown in FIG. 20 and Sequence 32 (HI2) is shown in FIG. 21.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11953) GTTTCCCTTGAGGCCAGGTCTTGTTAAGAAGAACAGAGAGCCCTGAGAGT ATTTCACGATGGTTACTTACCCCTTCTCCCTGGCAAAAGCAAAGCAGATT TTTCTCAGATCTTTACAGTGAGAATCTGACAGGATTCACAGAGGTAAAAC TGAGGTAAGTATTGAGGCCCCTCTCAGACTGAGCCTCCTTGGAGTTTTTT AACTCTCAAGCTAGTCTGCACTGAGCCTCCAGCAATTCCCCAATTACAGT TTAGTGTTCCTACTGATGTTGGCTCCAGCTGTGAAACTAGCTTCAGCTTC TGGCTTCTGTGCCTGGGCTCTGCTCCTGGTAAACTGTGATTCTCTGAAAA GCTGTGATTCTCTGTATCTATCTGTCTGTCTCTCTAGTTTTTAGGGCAGT GGTTTTTCCTGTGACCTCAATTCTCTGGTGGATCTAAGAAGAGTTACTGA TTTCAGTTTGCTTAGCTTTTTTTTTTCTTGTTGTGAGGATGGGAGTGACA ACTTCCAAGCTCTTTACATGTTGAACAGGAAACTGAAAGCCCCTTGGTGT TCCTTTGTAAATTCATCTTAAAAATATTTATCATAATTGAAAAGTGCTAA TATCAAATTTTCAGTCTGTTTATATTCCCCCTAAACTCAGATAAATATAC ATTTTATTTTGTGTGTCTGTGTGTGTGGGTTTTGTTGTTGTTTGTTTTTT GTGTTTTTTTTTAAGATATAGGGTCTTGCTCTGTCAAGGCTGGAGTGCAG TGGCACAATCGTACATCTCTGCAGCCTCGAACTCCTGGGAGAAAGTGATC CTCCCGCTTTAGCCTTCAGAGTAGCTACGACTACAGGCACTAACCACCAA GCCCAGCTAATTTTTAAAATTTTTTGTAGAGATGGGAGTTTCACTTTGTT GCCCAGGCTGGTCTCAAACTCTTGGCCTCAAGTGATCCTCCTGCCTCAGC CTCCCAAAATGTTGGAATCACAGATACTTTGTGTCTTGATTCTTGAAAGG AAAAAACAAAGATTTTTAATGCCTCTTATCTTGTACGCACTTTCCTTCCA AACAATACCCTTTTGCTGCCATTGTTCTCGTTATGAATAGCTTAAAGAAA AAGAAACAACTAAGGGTAGTAATAGGCCAGGAATCACTTACTGAATACTA GGTCTTCTTGTATAGTTTGATACCCTATAAATTGTGTGCATCTGATGCAT TTCACCTTCAAAAGGCTCAATGCTCTGTATTATTTAGTAGTAATCAAAAT TTCAAGTTTTACTTAACCTCCTGATTCACTGCCCAATTTCCTAATAAATA CGGGCTAAGGGTCAATGGGGTCATTTGCAAGTAATCTTGTAGTCTACTCA GAAAGTTCTGCAAAGTTAGAAAGTGATTAAATGACTGTTTGTTAAGATAT ACTTACATAGTAATAACCTAAATGCATTTGTTAAGTGGTTGTAGAGAGAG GGATTTAAAATTTTATCCTATATGAAATTTTCCTTTTTGGTGTCTGTTAT TTAATAGGATTGTTTGAATTAGGGGATACTATTTGGTGCCTTTGTAACTA TATGAAAATTAGTTGGTTGAATATTACTGCTTTCCATGTTCATATTTATA TTTGTATAGACATATATATATATACACATATACTACTTTCCTTTCCATTT TCATATTTATATTTGTGTATACACATATACATAAACATATATTTTATACA TTTTTGAAAAGGAAAATTAACTTAAGGGCATATTTAATGAATATTCAAAA ATTTTTTTGCTGATCAAATTATCATTCTGCTTTAAACTTTTGAAATGATC CAAAAAAATTTTAAATGACTTAGATTTACTGTTACAAAATGCTTGTCTTT TGATGTCACAAACATTATATACTATAATCACTGGCCAGAGATAATTGCTA TAAGTATAATGAAAAGGGAAATGATGGAAGAATCTCTGCAGCTATCCTCA TAAATGAGGGTGGGAACACGATGGGCAGTTCCAAAGTTGAAAATAGAGAA TATATGTGGATTTATATTAACATAATTGGTATTCTTGGATAGTTAAAAAT GGCTAAACTGTAGGAGAAGCCCGAGTAATTACTGTTAACAGAGGAATAAA TTTGAGGGCAATAATAATGATGATAGGCCAGGCACTGTGGCTCATGCCTG TAATCCCAGCACTTTGGGAACCCGAGGCGAGCGGACCACCTGAGGTCAGG AGTTCGAGAGCAGCCTGGCCAACATGGTGAAACCTCGTCTCTACTAAAAA TAGAAAAATTATCCGAGTGTGGTGGTGCGTGCCTGTAATCCCAGCTACTT GGGAGGCTGAGGCAGGAGAATCACTTGTACCTGGGAGGCGGAGTTGCAGT GAGCCGAAATCGCGCCACTGCGCTCCAGCCTGTGGGCCAGAGCGAGACTC CGCCTCAGAATAATAATAATGATAATAATAATAACGCCACCAACAATACT AAGAGCTAACATTTACTGAGTGCTTACTATGCACCAGATATTGTTCTAAG TATACATTTATTATCTCATTTAACCATCCATAATACTGTGGTATAGACAC TTTTATATCCATTTTATAAATAAGTAAACTGAGTTATGGAGAGATTAAAC GACTTGCCAGTAAGATTCAAAGCCTGTGTACAAGCTCACGCTTGATTCTG GAGCCAGTGTTCTTAACACAGTATCTTGAGAATGTTAAACTAAAAAGTTT TTAATTTACAGTATTCTTTCCACAATTAAAAAAGAAATTATGAGTAATTA TTTTTAGTTCTTTCTTCTCTTCAGGCATTTCCCATGGTTCTTTTCAAGAC ATAATACATATCATTTAGTGTTGTAGATCTGAAAAAACAAAAGTAGCGTG AAGATCAAAAATTTTCTAAAGAGACGGAGTCTCGCTACGTTCCCTAGGCT GGAACACCCAGGCTTCTCCAGCCTCACACCTCTGAGTAGCTGGAACCACC CTGTCCGCTAAGGTCAATGTTTAATCGTATCTTTGTAGGTCTACTGACCA GTTAAAAAGAGGTGCTGTATACATTGGTTGTTGTCTTGTCAGAGTTTGAT GCTTCTATATAGACCATTGTTTTTACATGCTAATACAATTGAAAGCCACT ACAGATATTTATATTTACAACCCAAAGCTAGGTTTTAACAAGAAACTCAT AAGGCAAAGGTGAGAAGTAAAATAATTTAGCGCCAAGTGGAGATATATGT GCAATGCTACTTTGTTGGGCTCAAAACATATTTTTCTTTTAGAAGACTGA CAGGCTTGAAGTTTATGCCTCCAAAGACAAAAGTGATTATGTTTTGTTTA GTAGCTTGCAAAGTTGCCAAAGCCATTTTTTCTACTCTTTCCCTGAAATT GGTTTATATGCTTATTAAAGTCATTTATACCTATTTGCAAATGCTTAACA TAGTTTCAGATTTTAAGATTTCCCTGCAACTTTATTTCCCTTGAAGTTTA CAGCAACAGGAGTTCATTTTTATTTTTAATTGCATTTATTCAGTAAGTAA ACTCCGCCACAGAAAAACTTAGTAGACAAGGTGAGTTCCCCTGTGCTCCG TGGCAAAGAGTGCGGTGGGTGACATTGACCCATGGTTAGGTAATCTGGTA AGGAAAGACCCCGTTGTAACACATCTGAGCAACGAGACCAAAGGAAGGGC TTGCTGCCACGAGGCGAAGTCTGCTTTTTTGAACAGAGAGCCCAGCAGAG TTGGGCGGCAATCGTGCCCAGCACTGAGGCCGAGGAGAAAGAGAGCAGGA GCATTACATTACTGCACCAAGAGTAGGAAAATATGATGCATGTTTGGGAC CAGGCAACCGAAATCCCTTCTCAGCAGCGCCTCCCAAAGCCGGGCACCGC CTTCCTTCGGAGAAGGCGCAGAGTCCCCAGACTCGGGCTGAGCCGCACCC CCATCTCCTTTCTCTTTCCTCCGCCGCTAAACACAGACGAGCACGTGAGC GTCGCAGCCCGTCCCAGCTGTGCCTCAGCTGACCGCCTCCTGATTGGCTG AGAGCGGCGTGGGCTGGGGTGGGGACTTGCCGCCTGCGTCGCTCGCCATT GGATCTCGAGGAACCCGCCTCCACCTCAGGTGAGGCGGGCTTGCGGGAGC GCGCGCCGGCCTGGGCAGGCGAGCGGGCGCGCTCCCGCCCCCTCTCCCCT CCCCGCGCGCCCGAGCGCGCCTCCGCCCTTGCCCGCCCCCTGACGCTGCC TCAGCTCCTCAGTGCACAGTGCTGCCTCGTCTGAGGGGACAGGAGGATCA CCCTCTTCGTCGCTTCGGCCAGTGTGTCGGGCTGGGCCCTGACAAGCCAC CTGAGGAGAGGCTCGGAGCCGGGCCCGGACCCCGGCGATTGCCGCCCGCT TCTCTCTAGTCTCACGAGGGGTTTCCCGCCTCGCACCCCCACCTCTGGAC TTGCCTTTCCTTCTCTTCTCCGCGTGTGGAGGGAGCCAGCGCTTAGGCCG GAGCGAGCCTGGGGGCCGCCCGCCGTGAAGACATCGCGGGGACCGATTCA CC ATG

5) IL-8. IL-8 is a member of the CXC chemokine family. IL-8 is a chemokine produced by macrophages, immune and epithelial cells and is an important mediator of immune reaction in the innate immune system (reviewed in Waugh and Wilson, 2008; Clin Cancer Res 14; 6735). While neutrophil granulocytes are the primary target cells of IL-8, there is a relative wide range of cells (endothelial cells, macrophages, mast cells, and keratinocytes) respond to IL-8. IL-8, also known as neutrophil chemotactic factor, has two primary functions. It induces chemotaxis in target cells, primarily neutrophils but also other granulocytes, causing them to migrate toward the site of infection. IL-8 also induces phagocytosis once they have arrived. IL-8 is also known to be a potent promoter of angiogenesis. In target cells, IL-8 induces a series of physiological responses required for migration and phagocytosis, such as increase of intracellular Ca2+, exocytosis (e.g. histamine release), and respiratory burst.

IL-8 can be secreted by any cells with toll-like receptors that are involved in the innate immune response. Generally, macrophages see the antigen first, and thus are first to release IL-8 to recruit other cells. Both monomer and homodimer forms of IL-8 have been reported to be potent inducers of the chemokines CXCR1 and CXCR2. The homodimer is more potent, but methylation of Leu25 can block activity of the dimers. IL-8 is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection. IL-8 is implicated in gingivitis, psoriasis and increased oxidant stress thereby enhancing the recruitment of inflammatory cells to the site of local inflammation.

Protein: IL-8 Gene: IL-8 (Homo sapiens, chromosome 4, 74606223-74609433 [NCBI Reference Sequence: NC_(—)000004.11]; start site location: 74606376; strand: positive)

Gene Identification GeneID 3576 HGNC 6025 HPRD 00909 MIM 146930

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 1314 IL8-1 ACGTCCCATTCGGCTCCTGAGCCA 2868 1331 IL8-3 GACGTTGACGAAGTCTATCACCCAA 2939 1341 ACGGAGTATGACGAAAGTTTTC 257 1342 GAGCGAGACTCCCGTCTAAA 3259

Target Shift Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 1314 ACGTCCCATTCGGCTCCTGAGCCA 2868 1315 CGTCCCATTCGGCTCCTGAG 2869 1316 GTCCCATTCGGCTCCTGAGC 2870 1317 TCCCATTCGGCTCCTGAGCC 2871 1318 CCCATTCGGCTCCTGAGCCA 2872 1319 CCATTCGGCTCCTGAGCCAT 2873 1320 CATTCGGCTCCTGAGCCATA 2874 1321 ATTCGGCTCCTGAGCCATAA 2875 1322 TTCGGCTCCTGAGCCATAAG 2876 1323 TCGGCTCCTGAGCCATAAGA 2877 1324 CGGCTCCTGAGCCATAAGAA 2878 1325 TACGTCCCATTCGGCTCCTG 2867 1326 TTACGTCCCATTCGGCTCCT 2866 1327 TTTACGTCCCATTCGGCTCC 2865 1328 ATTTACGTCCCATTCGGCTC 2864 1329 TATTTACGTCCCATTCGGCT 2863 1330 TTATTTACGTCCCATTCGGC 2862 1331 GACGTTGACGAAGTCTATCACCCAA 2939 1332 ACGTTGACGAAGTCTATCAC 2940 1333 CGTTGACGAAGTCTATCACC 2941 1334 GTTGACGAAGTCTATCACCC 2942 1335 TTGACGAAGTCTATCACCCA 2943 1336 TGACGAAGTCTATCACCCAA 2944 1337 GACGAAGTCTATCACCCAAG 2945 1338 ACGAAGTCTATCACCCAAGA 2946 1339 CGAAGTCTATCACCCAAGAA 2947 1340 AGACGTTGACGAAGTCTATC 2938 1341 ACGGAGTATGACGAAAGTTTTC 257 1342 GAGCGAGACTCCCGTCTAAA 3259 1343 AGCGAGACTCCCGTCTAAAA 3260 1344 GCGAGACTCCCGTCTAAAAA 3261 1345 CGAGACTCCCGTCTAAAAAA 3262 1346 GAGACTCCCGTCTAAAAAAG 3263 1347 AGAGCGAGACTCCCGTCTAA 3258 1348 AAGAGCGAGACTCCCGTCTA 3257 1349 AAAGAGCGAGACTCCCGTCT 3256 1350 GAAAGAGCGAGACTCCCGTC 3255 1351 TGAAAGAGCGAGACTCCCGT 3254 1352 GTGAAAGAGCGAGACTCCCG 3253 1353 GGTGAAAGAGCGAGACTCCC 3252

Hot Zones (Relative upstream location to gene start site)  1-300 2650-3300 4800-5000

Examples

In FIG. 22, IL8-1 (41) and IL8-3 (42), both at 10 μM, demonstrated statistically significant (P<0.05) inhibition compared to the untreated control values in MDA-MB-231 (human breast cell line). The negative control did not produce a statistically significant difference compared to the untreated control. The IL-8 sequences IL8-1 (41) and IL8-3 (42) fit the independent and dependent DNAi motif claims.

In FIG. 23, IL8-1 (41) and IL8-3 (42), both at 10 μM, demonstrated statistically significant (P<0.05) inhibition compared to the untreated control values in DU145 (human prostate cell line). The negative control did not produce a statistically significant difference compared to the untreated control. The IL-8 sequences IL8-1 (41) and IL8-3 (42) fit the independent and dependent DNAi motif claims.

The secondary structures for IL8-1 and IL8-3 are shown in FIGS. 24 and 25. Sequence 41 (IL8-1) is shown in FIG. 24 and Sequence 42 (IL8-3) is shown in FIG. 25.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11954) GGCATTAAAAAAGAAAGCTTATATAGTGGAAGAAAATAAAGCATCTAGAC ATAAGCTTTAAGAGATCTATTGTGTTAATACAGCTTTACTTTTTGAGTGG TAAGCTTTTAAAAAGAAATGTGGTGCTCTAACTCCAGGAAAAGATAAGGG TGACTGAAGTGATAGTCTAGAGGAAAAAGATGCAGACATTTACTGAGTAC CTCCAATGTGCCAGGTGCCATTCTGGGCATTTTCATTATGTTTCCTCATT TAATTCTCATGGTGATCCTTTGGAACTGTGTTATTCTCATTTTTACAGAT GAGGTAACTGAGAGACAGTCAGATTAAAGAACTGCCTATGATTGTTTGGC TAATAATAAGTGGAGGGGTGAGGCTTGAAGGCAGGTTTGTCTTATTCCAA CACCCATACATACCCTTAAATTTAAGTTATTCTGACTTGTGTTGCTCAAA TCCAATGTGTTCAGCTGTTTGCTTCTCCAATTACCAAGATTTTTCTTTAA AAGGTAGGACACTTTTGGCAACACGAACCAACTTTGCTCAGTATTGTTAT AAACTGTTAACTGGAGACATTTGAATTTGGAGATGGAACTGAAATGGTCT TGCGGTACTAGAGAAGATCAAGTTATCACATAAACAAAGTACAGAGCTGA GAACATATTTTAAATCTTTCCACTAACTCTGACTTTTATTGACTAAAATT TTAGTGGGCAGTATGTTTATGTTTATGACTCTTAACATTAACAACATCGT AAGTCAAACTCACTAATATATGTTAAGCATTCTGTTTATGATTCTTTTAA CCTAGAGGATTGTTGAGCTGGGACTAATTTCCTCAAATGGGAAAAAAACC CAGGTGAGAGCTGAGACTGCTCCTGAGACTGAGAAAGGCAGCTCTGACGG GATCTCAGATTTTAGCAGCAGGAGTTGAACAATGGGCATAGAATCAGCTT GCCCAAGATCTCCTGATTAATAAACCATGGAACAAGATTTAAACCCAAGT TCATTTCATTTCAAAGCTCATACCACATTTTGCCCACCATATTTTGCTTT GTTATATGACTACAACTTAGTTCAGGCTTACAAAAAAGTCCTAATTCTAA AATTCCTATGGCGTGGGTGGGAGGGGATTTAGATGATTTTGCATAGGCAA GAAACACCCAGTTTCATGGAGTTTGATGGAAGAGTTATGTACTAATATGG GAAAAGTAGAGGCCATCTTTGTCTTTGTTCTTTCTTTTTTAGACGGGAGT CTCGCTCTTTCACCCAGGCTGGAGTGCAGTGGCGCTATCTCGGCTCACTG CAAGCTCCGCCTCCTGGGTTCACGCCATTCTCCTGCCTCAGCCTCCTGAG TAGCTGGGACTACAGGCGCCCGCCACCGCTCCCGGCTAATTTTTTATATT TTCAGTAGAGACGGGGTTTCACCGTGTTAGCTAAGATGGTTTGGATCTCC TGAACTCGTGATCCGCCCGCCTCGGACTCCCAAAGTGCTGGGATTACAGG CTTGAGCCACCGCGCCCGGCCGTCTTTGTTCTTTCTTGAACTCTTCCTTT TCTTGGGTGATAGACTTCGTCAACGTCTAATGAGGATATCTAGGTGCTAG TCTCTGCTCATCAAATGATTCTTATGGCTCAGGAGCCGAATGGGACGTAA ATAAACAGTTAAGTCTCATGAACTCACTTTGCATTCATCTCTAGAAGATG ACAAAACATTTGTATTTATGTGTAGCGTGGCACTTTAGTTAAACTTTGTA CCCCACTTTGCTCTATTTTAAAGCAGAATATCCTTAAAAAGGATACTTAG TCCTGCTTTTTTTTTTCCGCCTAAGCCCATTTAGTCCTTCTACTCATTAT GCAAGGACTCAAATGGTTATCTTTACAGAAGTGAGACAAGATAGAATCAA TGCTCTTGTAGTCACTTCATCTTTGTCCATTCCCACTTCTGATGGAGAGG GTTCTAGGACATAATGCACTGAAGGTTACATTGTGAGAGATGAACAACAT TTGCAAAAGAGGTCTTTTTGCCTTGGAAAGGCTTCATTCTTAAAAAAAAA TGTGAGCATCAAGGTTAAGTAGACCTCATTAGCTCAAACTTTAAGGATGA TATCAGGATAAAGTTGGGCCCATGAGAAGAGAATGAGAGGGAGATATAGT GACATGAAAATAAGGAGGAAAACGAGGTGTCTATGTAAGTTGGGCTCACC ATAAATACAAAGGCAACCGTTAGGGAAAAGCAAAGAAGTCTTTGCACATC CTCAGAACTCTGAATGTCTTAGTGATGCTGTATGAGTGAGTCTTAATGAT AGTGAACTGAATCAGTCAAGCCAGGTTGTGTCCATATGAGAATGTGTCTT TGCTAAACATGCCAACATCACTGAAGCAAAGAAACTTGGAGTTTTCTTTA AGATATAGGTCTTTTTTACCTATCCGGCCCAAGCTTTCTCTTCTTGTCAC TCCATGCACTGTGTTCCGTATGCTAAATAGTTTGAGAAACCCAAATGGGC CATGTTCGCCTACATTTCATTGTCCTGTACTTCCTGTCCTGTACTAGCAA AGCAGTCCCATTGGTCTTTCTTCTCCTCATTAACAATAAAGGTAACACTT TTGATGTTGTTTCTTCAGAAAACCTTCATTCATCAAAACTGCCTCAAAGA TCATGTTTGTTTGATTCCAGAACTTCCTGTAATTACCTGTTATTGTAACA CTCATCACTGTATTTTACTTACTTGTGTAACTAATTTTCCATATTCTGCA CTAGACAACAAAGTCCTTTAAGTCAGGTACTATATCTATTTACATAGCAT TCACATCTCCTACAATAAGGGACATTAGCAGATAAACAACACATATTAAA TGAATAATGAAGTTTCTGAAATACTACAGTTGAAAACTATAGGAGCTACA TTATATAGAATAAACATTTACTTTGCTATAGAATTCAGTGTAACCCAGGC ATTATTTTATCCTCAAGTCTTAGGTTGGTTGGAGAAAGATAACAAAAAGA AACATGATTGTGCAGAAACAGACAAACCTTTTTGGAAAGCATTTGAAAAT GGCATTCCCCCTCCACAGTGTGTTCACAGTGTGGGCAAATTCACTGCTCT GTCGTACTTTCTGAAAATGAAGAACTGTTACACCAAGGTGAATTATTTAT AAATTATGTACTTGCCCAGAAGCGAACAGACTTTTACTATCATAAGAACC CTTCCTTGGTGCTCTTTATCTACAGAATCCAAGACCTTTCAAGAAAGGTC TTGGATTCTTTTCTTCAGGACACTAGGACATAAAGCCACCTTTTTATGAT TTGTTGAAATTTCTCACTCCATCCCTTTTGCTAGTGATCATGGGTCCTCA GAGGTCAGACTTGGTGTCCTTGGATAAAGAGCATGAAGCAACAGTGGCTG AACCAGAGTTGGAACCCAGATGCTCTTTCCACTAAGCATACAACTTTCCA TTAGATAACACCTCCCTCCCACCCCAACCAAGCAGCTCCAGTGCACCACT TTCTGGAGCATAAACATACCTTAACTTTACAACTTGAGTGGCCTTGAATA CTGTTCCTATCTGGAATGTGCTGTTCTCTTTCATCTTCCTCTATTGAAGC CCTCCTATTCCTCAATGCCTTGCTCCAACTGCCTTTGGAAGATTCTGCTC TTATGCCTCCACTGGAATTAATGTCTTAGTACCACTTGTCTATTCTGCTA TATAGTCAGTCCTTACATTGCTTTCTTCTTCTGATAGACCAAACTCTTTA AGGACAAGTACCTAGTCTTATCTATTTCTAGATCCCCCACATTACTCAGA AAGTTACTCCATAAATGTTTGTGGAACTGATTTCTATGTGAAGCACATGT GCCCCTTCACTCTGTTAACATGCATTAGAAAACTAAATCTTTTGAAAAGT TGTAGTATGCCCCCTAAGAGCAGTAACAGTTCCTAGAAACTCTCTAAAAT GCTTAGAAAAAGATTTATTTTAAATTACCTCCCCAATAAAATGATTGGCT GGCTTATCTTCACCATCATGATAGCATCTGTAATTAACTGAAAAAAAATA ATTATGCCATTAAAAGAAAATCATCCATGATCTTGTTCTAACACCTGCCA CTCTAGTACTATATCTGTCACATGGTACTATGATAAAGTTATCTAGAAAT AAAAAAGCATACAATTGATAATTCACCAAATTGTGGAGCTTCAGTATTTT AAATGTATATTAAAATTAAATTATTTTAAAGATCAAAGAAAACTTTCGTC ATACTCCGTATTTGATAAGGAACAAATAGGAAGTGTGATGACTCAGGTTT GCCCTGAGGGGATGGGCCATCAGTTGCAAATCGTGGAATTTCCTCTGACA TAATGAAAAGATGAGGGTGCATAAGTTCTCTAGTAGGGTGATGATATAAA AAGCCACCGGAGCACTCCATAAGGCACAAACTTTCAGAGACAGCAGAGCA CACAAGCTTCTAGGACAAGAGCCAGGAAGAAACCACCGGAAGGAACCATC TCACTGTGTGTAAAC ATG

6) KRAS or GTPase KRas also known as V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog and KRAS, is a protein that in humans is encoded by the KRAS gene (McGrath et al. Nature 1983; 304 (5926): 501-6, Popescu et al., Somat. Cell Mol. Genet. 1985; 11 (2): 149-55) and is usually tethered to cell membranes because by its C-terminal isoprenyl group. The protein product of the normal KRAS gene performs an essential function in normal tissue signaling. A single amino acid substitution resulting from a particular single nucleotide substitution in genomic DNA, is responsible for the activating mutation. Once on, it recruits and activates C-RAF and PI3Kinase necessary for to propagate growth factor and other receptor signals. The transformed protein that results is implicated in various malignancies, including leukemias, lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas and colorectal carcinoma (Kranenburg, Biochim. Biophys. Acta 2005; 1756 (2): 81-2; Burmer and Loeb, Proc. Natl. Acad. Sci. U.S.A. 86 (7): 2403-7, Tam et al, Clin. Cancer Res. 12 (5): 1647-53, Almoguera et al, Cell 53 (4): 549-54). Several germline KRAS mutations have been found to be associated with Noonan syndrome (Gelb and Tartaglia, Human Molecular Genetics, 2006; 15 (2): R220-226).

Protein: KRAS Gene: KRAS (Homo sapiens, chromosome 12, 25358180-25403854 [NCBI Reference Sequence: NC_(—)000012.11]; start site location: 25398318; strand: negative)

Gene Identification GeneID 3845 HGNC 6407 HPRD 01817 MIM 190070

Targeted Sequences Relative upstream Sequence Design location to gene start ID No: ID Sequence (5′-3′) site 1354 KR1 CCCGGAGCGGGACCGGACCGCGG 5923 1435 KR2 GCCGGACCCACGCGGCGGCCCGCC 5856 1516 KR0525 AGTCTCCCCTTCCCGGAGACT 10265 1535 GCCGGGCCGGCTGGAGAGCGGGTC 5803 1538 TCGCCCCTCCTCCGAGACTTTC 6626 1584 GCACCCCGCCACCCTCAGGGTCGGC 6029 1633 GAGCCGCCGCCACCTTCGCCGCCGC 5475 1697 CGGCATAGTTCCCCGCCTTAC 2002 1730 KR16 CGGCCCGAGCCTCCGTGACGAGTGC 146348 1767 KR17 CTGGGAGGGGATCCCTCACCGAGAG 3328

Target Shift Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 1354 CCCGGAGCGGGACCGGACCGCGG 5923 1355 CCGGAGCGGGACCGGACCGC 5924 1356 CGGAGCGGGACCGGACCGCG 5925 1357 ACCCGGAGCGGGACCGGACC 5922 1358 GACCCGGAGCGGGACCGGAC 5921 1359 TGACCCGGAGCGGGACCGGA 5920 1360 CTGACCCGGAGCGGGACCGG 5919 1361 TCTGACCCGGAGCGGGACCG 5918 1362 TTCTGACCCGGAGCGGGACC 5917 1363 ATTCTGACCCGGAGCGGGAC 5916 1364 AATTCTGACCCGGAGCGGGA 5915 1365 CAATTCTGACCCGGAGCGGG 5914 1366 CCAATTCTGACCCGGAGCGG 5913 1367 GCCAATTCTGACCCGGAGCG 5912 1368 CGCCAATTCTGACCCGGAGC 5911 1369 CCGCCAATTCTGACCCGGAG 5910 1370 GCCGCCAATTCTGACCCGGA 5909 1371 AGCCGCCAATTCTGACCCGG 5908 1372 CAGCCGCCAATTCTGACCCG 5907 1373 GCAGCCGCCAATTCTGACCC 5906 1374 CGCAGCCGCCAATTCTGACC 5905 1375 CCGCAGCCGCCAATTCTGAC 5904 1376 CCCGCAGCCGCCAATTCTGA 5903 1377 CCCCGCAGCCGCCAATTCTG 5902 1378 TCCCCGCAGCCGCCAATTCT 5901 1379 GTCCCCGCAGCCGCCAATTC 5900 1380 TGTCCCCGCAGCCGCCAATT 5899 1381 CTGTCCCCGCAGCCGCCAAT 5898 1382 GCTGTCCCCGCAGCCGCCAA 5897 1383 GGCTGTCCCCGCAGCCGCCA 5896 1384 AGGCTGTCCCCGCAGCCGCC 5895 1385 AAGGCTGTCCCCGCAGCCGC 5894 1386 CAAGGCTGTCCCCGCAGCCG 5893 1387 GCAAGGCTGTCCCCGCAGCC 5892 1388 CGCAAGGCTGTCCCCGCAGC 5891 1389 CCGCAAGGCTGTCCCCGCAG 5890 1390 GCCGCAAGGCTGTCCCCGCA 5889 1391 AGCCGCAAGGCTGTCCCCGC 5888 1392 TAGCCGCAAGGCTGTCCCCG 5887 1393 CTAGCCGCAAGGCTGTCCCC 5886 1394 CCTAGCCGCAAGGCTGTCCC 5885 1395 GCCTAGCCGCAAGGCTGTCC 5884 1396 TGCCTAGCCGCAAGGCTGTC 5883 1397 CTGCCTAGCCGCAAGGCTGT 5882 1398 CCTGCCTAGCCGCAAGGCTG 5881 1399 CCCTGCCTAGCCGCAAGGCT 5880 1400 CCCCTGCCTAGCCGCAAGGC 5879 1401 CCCCCTGCCTAGCCGCAAGG 5878 1402 GCCCCCTGCCTAGCCGCAAG 5877 1403 CGCCCCCTGCCTAGCCGCAA 5876 1404 CCGCCCCCTGCCTAGCCGCA 5875 1405 CCCGCCCCCTGCCTAGCCGC 5874 1406 GCCCGCCCCCTGCCTAGCCG 5873 1407 GGCCCGCCCCCTGCCTAGCC 5872 1408 CGGCCCGCCCCCTGCCTAGC 5871 1409 GCGGCCCGCCCCCTGCCTAG 5870 1410 GGCGGCCCGCCCCCTGCCTA 5869 1411 CGGCGGCCCGCCCCCTGCCT 5868 1412 GCGGCGGCCCGCCCCCTGCC 5867 1413 CGCGGCGGCCCGCCCCCTGC 5866 1414 ACGCGGCGGCCCGCCCCCTG 5865 1415 CACGCGGCGGCCCGCCCCCT 5864 1416 CCACGCGGCGGCCCGCCCCC 5863 1417 CCCACGCGGCGGCCCGCCCC 5862 1418 ACCCACGCGGCGGCCCGCCC 5861 1419 GACCCACGCGGCGGCCCGCC 5860 1420 GGACCCACGCGGCGGCCCGC 5859 1421 CGGACCCACGCGGCGGCCCG 5858 1422 CCGGACCCACGCGGCGGCCC 5857 1423 GCCGGACCCACGCGGCGGCC 5856 1424 TGCCGGACCCACGCGGCGGC 5855 1425 CTGCCGGACCCACGCGGCGG 5854 1426 ACTGCCGGACCCACGCGGCG 5853 1427 GACTGCCGGACCCACGCGGC 5852 1428 GGACTGCCGGACCCACGCGG 5851 1429 GGGACTGCCGGACCCACGCG 5850 1430 AGGGACTGCCGGACCCACGC 5849 1431 GAGGGACTGCCGGACCCACG 5848 1432 GGAGGGACTGCCGGACCCAC 5847 1433 AGGAGGGACTGCCGGACCCA 5846 1434 GAGGAGGGACTGCCGGACCC 5845 1435 GCCGGACCCACGCGGCGGCCCGCC 5856 1436 CCGGACCCACGCGGCGGCCC 5857 1437 CGGACCCACGCGGCGGCCCG 5858 1438 GGACCCACGCGGCGGCCCGC 5859 1439 GACCCACGCGGCGGCCCGCC 5860 1440 ACCCACGCGGCGGCCCGCCC 5861 1441 CCCACGCGGCGGCCCGCCCC 5862 1442 CCACGCGGCGGCCCGCCCCC 5863 1443 CACGCGGCGGCCCGCCCCCT 5864 1444 ACGCGGCGGCCCGCCCCCTG 5865 1445 CGCGGCGGCCCGCCCCCTGC 5866 1446 GCGGCGGCCCGCCCCCTGCC 5867 1447 CGGCGGCCCGCCCCCTGCCT 5868 1448 GGCGGCCCGCCCCCTGCCTA 5869 1449 GCGGCCCGCCCCCTGCCTAG 5870 1450 CGGCCCGCCCCCTGCCTAGC 5871 1451 GGCCCGCCCCCTGCCTAGCC 5872 1452 GCCCGCCCCCTGCCTAGCCG 5873 1453 CCCGCCCCCTGCCTAGCCGC 5874 1454 CCGCCCCCTGCCTAGCCGCA 5875 1455 CGCCCCCTGCCTAGCCGCAA 5876 1456 GCCCCCTGCCTAGCCGCAAG 5877 1457 CCCCCTGCCTAGCCGCAAGG 5878 1458 CCCCTGCCTAGCCGCAAGGC 5879 1459 CCCTGCCTAGCCGCAAGGCT 5880 1460 CCTGCCTAGCCGCAAGGCTG 5881 1461 CTGCCTAGCCGCAAGGCTGT 5882 1462 TGCCTAGCCGCAAGGCTGTC 5883 1463 GCCTAGCCGCAAGGCTGTCC 5884 1464 CCTAGCCGCAAGGCTGTCCC 5885 1465 CTAGCCGCAAGGCTGTCCCC 5886 1466 TAGCCGCAAGGCTGTCCCCG 5887 1467 AGCCGCAAGGCTGTCCCCGC 5888 1468 GCCGCAAGGCTGTCCCCGCA 5889 1469 CCGCAAGGCTGTCCCCGCAG 5890 1470 CGCAAGGCTGTCCCCGCAGC 5891 1471 GCAAGGCTGTCCCCGCAGCC 5892 1472 CAAGGCTGTCCCCGCAGCCG 5893 1473 AAGGCTGTCCCCGCAGCCGC 5894 1474 AGGCTGTCCCCGCAGCCGCC 5895 1475 GGCTGTCCCCGCAGCCGCCA 5896 1476 GCTGTCCCCGCAGCCGCCAA 5897 1477 CTGTCCCCGCAGCCGCCAAT 5898 1478 TGTCCCCGCAGCCGCCAATT 5899 1479 GTCCCCGCAGCCGCCAATTC 5900 1480 TCCCCGCAGCCGCCAATTCT 5901 1481 CCCCGCAGCCGCCAATTCTG 5902 1482 CCCGCAGCCGCCAATTCTGA 5903 1483 CCGCAGCCGCCAATTCTGAC 5904 1484 CGCAGCCGCCAATTCTGACC 5905 1485 GCAGCCGCCAATTCTGACCC 5906 1486 CAGCCGCCAATTCTGACCCG 5907 1487 AGCCGCCAATTCTGACCCGG 5908 1488 GCCGCCAATTCTGACCCGGA 5909 1489 CCGCCAATTCTGACCCGGAG 5910 1490 CGCCAATTCTGACCCGGAGC 5911 1491 GCCAATTCTGACCCGGAGCG 5912 1492 CCAATTCTGACCCGGAGCGG 5913 1493 CAATTCTGACCCGGAGCGGG 5914 1494 AATTCTGACCCGGAGCGGGA 5915 1495 ATTCTGACCCGGAGCGGGAC 5916 1496 TTCTGACCCGGAGCGGGACC 5917 1497 TCTGACCCGGAGCGGGACCG 5918 1498 CTGACCCGGAGCGGGACCGG 5919 1499 TGACCCGGAGCGGGACCGGA 5920 1500 GACCCGGAGCGGGACCGGAC 5921 1501 ACCCGGAGCGGGACCGGACC 5922 1502 CCCGGAGCGGGACCGGACCG 5923 1503 CCGGAGCGGGACCGGACCGC 5924 1504 CGGAGCGGGACCGGACCGCG 5925 1505 TGCCGGACCCACGCGGCGGC 5855 1506 CTGCCGGACCCACGCGGCGG 5854 1507 ACTGCCGGACCCACGCGGCG 5853 1508 GACTGCCGGACCCACGCGGC 5852 1509 GGACTGCCGGACCCACGCGG 5851 1510 GGGACTGCCGGACCCACGCG 5850 1511 AGGGACTGCCGGACCCACGC 5849 1512 GAGGGACTGCCGGACCCACG 5848 1513 GGAGGGACTGCCGGACCCAC 5847 1514 AGGAGGGACTGCCGGACCCA 5846 1515 GAGGAGGGACTGCCGGACCC 5845 1516 AGTCTCCCCTTCCCGGAGACT 10265 1517 GTCTCCCCTTCCCGGAGACT 10266 1518 TCTCCCCTTCCCGGAGACTT 10267 1519 CTCCCCTTCCCGGAGACTTA 10268 1520 TCCCCTTCCCGGAGACTTAA 10269 1521 CCCCTTCCCGGAGACTTAAT 10270 1522 CCCTTCCCGGAGACTTAATC 10271 1523 CCTTCCCGGAGACTTAATCT 10272 1524 CTTCCCGGAGACTTAATCTT 10273 1525 TTCCCGGAGACTTAATCTTG 10274 1526 TCCCGGAGACTTAATCTTGC 10275 1527 CCCGGAGACTTAATCTTGCT 10276 1528 CCGGAGACTTAATCTTGCTT 10277 1529 CGGAGACTTAATCTTGCTTC 10278 1530 AAGTCTCCCCTTCCCGGAGA 10264 1531 TAAGTCTCCCCTTCCCGGAG 10263 1532 TTAAGTCTCCCCTTCCCGGA 10262 1533 GTTAAGTCTCCCCTTCCCGG 10261 1534 AGTTAAGTCTCCCCTTCCCG 10260 1535 GCCGGGCCGGCTGGAGAGCGGGTC 5803 1536 CCGGGCCGGCTGGAGAGCGG 5804 1537 AGCCGGGCCGGCTGGAGAGC 5802 1538 TCGCCCCTCCTCCGAGACTTTC 6626 1539 CGCCCCTCCTCCGAGACTTT 6627 1540 GCCCCTCCTCCGAGACTTTC 6628 1541 CCCCTCCTCCGAGACTTTCA 6629 1542 CCCTCCTCCGAGACTTTCAG 6630 1543 CCTCCTCCGAGACTTTCAGT 6631 1544 CTCCTCCGAGACTTTCAGTT 6632 1545 TCCTCCGAGACTTTCAGTTC 6633 1546 CCTCCGAGACTTTCAGTTCC 6634 1547 CTCCGAGACTTTCAGTTCCA 6635 1548 TCCGAGACTTTCAGTTCCAT 6636 1549 CCGAGACTTTCAGTTCCATT 6637 1550 CGAGACTTTCAGTTCCATTC 6638 1551 ATCGCCCCTCCTCCGAGACT 6625 1552 GATCGCCCCTCCTCCGAGAC 6624 1553 GGATCGCCCCTCCTCCGAGA 6623 1554 AGGATCGCCCCTCCTCCGAG 6622 1555 TAGGATCGCCCCTCCTCCGA 6621 1556 ATAGGATCGCCCCTCCTCCG 6620 1557 GATAGGATCGCCCCTCCTCC 6619 1558 TGATAGGATCGCCCCTCCTC 6618 1559 CTGATAGGATCGCCCCTCCT 6617 1560 CCTGATAGGATCGCCCCTCC 6616 1561 ACCTGATAGGATCGCCCCTC 6615 1562 TACCTGATAGGATCGCCCCT 6614 1563 GTACCTGATAGGATCGCCCC 6613 1564 TGTACCTGATAGGATCGCCC 6612 1565 CTGTACCTGATAGGATCGCC 6611 1566 CCTGTACCTGATAGGATCGC 6610 1567 GCCTGTACCTGATAGGATCG 6609 1568 CGCCTGTACCTGATAGGATC 6608 1569 GCGCCTGTACCTGATAGGAT 6607 1570 AGCGCCTGTACCTGATAGGA 6606 1571 CAGCGCCTGTACCTGATAGG 6605 1572 GCAGCGCCTGTACCTGATAG 6604 1573 AGCAGCGCCTGTACCTGATA 6603 1574 AAGCAGCGCCTGTACCTGAT 6602 1575 AAAGCAGCGCCTGTACCTGA 6601 1576 AAAAGCAGCGCCTGTACCTG 6600 1577 GAAAAGCAGCGCCTGTACCT 6599 1578 GGAAAAGCAGCGCCTGTACC 6598 1579 TGGAAAAGCAGCGCCTGTAC 6597 1580 CTGGAAAAGCAGCGCCTGTA 6596 1581 GCTGGAAAAGCAGCGCCTGT 6595 1582 GGCTGGAAAAGCAGCGCCTG 6594 1583 GGGCTGGAAAAGCAGCGCCT 6593 1584 GCACCCCGCCACCCTCAGGGTCGGC 6029 1585 CACCCCGCCACCCTCAGGGT 6030 1586 ACCCCGCCACCCTCAGGGTC 6031 1587 CCCCGCCACCCTCAGGGTCG 6032 1588 CCCGCCACCCTCAGGGTCGG 6033 1589 CCGCCACCCTCAGGGTCGGC 6034 1590 CGCCACCCTCAGGGTCGGCC 6035 1591 GCCACCCTCAGGGTCGGCCT 6036 1592 CCACCCTCAGGGTCGGCCTA 6037 1593 CACCCTCAGGGTCGGCCTAT 6038 1594 ACCCTCAGGGTCGGCCTATA 6039 1595 CCCTCAGGGTCGGCCTATAC 6040 1596 CCTCAGGGTCGGCCTATACT 6041 1597 CTCAGGGTCGGCCTATACTG 6042 1598 TCAGGGTCGGCCTATACTGG 6043 1599 CAGGGTCGGCCTATACTGGC 6044 1600 AGGGTCGGCCTATACTGGCG 6045 1601 GGGTCGGCCTATACTGGCGC 6046 1602 GGTCGGCCTATACTGGCGCG 6047 1603 GTCGGCCTATACTGGCGCGC 6048 1604 TCGGCCTATACTGGCGCGCA 6049 1605 CGGCCTATACTGGCGCGCAT 6050 1606 GGCCTATACTGGCGCGCATC 6051 1607 GCCTATACTGGCGCGCATCC 6052 1608 CCTATACTGGCGCGCATCCA 6053 1609 CTATACTGGCGCGCATCCAT 6054 1610 TATACTGGCGCGCATCCATT 6055 1611 ATACTGGCGCGCATCCATTT 6056 1612 TACTGGCGCGCATCCATTTA 6057 1613 ACTGGCGCGCATCCATTTAC 6058 1614 CTGGCGCGCATCCATTTACT 6059 1615 TGGCGCGCATCCATTTACTA 6060 1616 GGCGCGCATCCATTTACTAT 6061 1617 GCGCGCATCCATTTACTATC 6062 1618 CGCGCATCCATTTACTATCA 6063 1619 AGCACCCCGCCACCCTCAGG 6028 1620 GAGCACCCCGCCACCCTCAG 6027 1621 AGAGCACCCCGCCACCCTCA 6026 1622 AAGAGCACCCCGCCACCCTC 6025 1623 GAAGAGCACCCCGCCACCCT 6024 1624 CGAAGAGCACCCCGCCACCC 6023 1625 GCGAAGAGCACCCCGCCACC 6022 1626 TGCGAAGAGCACCCCGCCAC 6021 1627 CTGCGAAGAGCACCCCGCCA 6020 1628 GCTGCGAAGAGCACCCCGCC 6019 1629 AGCTGCGAAGAGCACCCCGC 6018 1630 AAGCTGCGAAGAGCACCCCG 6017 1631 GAAGCTGCGAAGAGCACCCC 6016 1632 AGAAGCTGCGAAGAGCACCC 6015 1633 GAGCCGCCGCCACCTTCGCCGCCGC 5475 1634 AGCCGCCGCCACCTTCGCCG 5476 1635 GCCGCCGCCACCTTCGCCGC 5477 1636 CCGCCGCCACCTTCGCCGCC 5478 1637 CGCCGCCACCTTCGCCGCCG 5479 1638 GCCGCCACCTTCGCCGCCGC 5480 1639 CCGCCACCTTCGCCGCCGCC 5481 1640 CGCCACCTTCGCCGCCGCCA 5482 1641 GCCACCTTCGCCGCCGCCAC 5483 1642 CCACCTTCGCCGCCGCCACT 5484 1643 CACCTTCGCCGCCGCCACTG 5485 1644 ACCTTCGCCGCCGCCACTGC 5486 1645 CCTTCGCCGCCGCCACTGCC 5487 1646 CTTCGCCGCCGCCACTGCCG 5488 1647 TTCGCCGCCGCCACTGCCGC 5489 1648 TCGCCGCCGCCACTGCCGCC 5490 1649 CGCCGCCGCCACTGCCGCCG 5491 1650 GCCGCCGCCACTGCCGCCGC 5492 1651 CCGCCGCCACTGCCGCCGCC 5493 1652 CGCCGCCACTGCCGCCGCCG 5494 1653 GCCGCCACTGCCGCCGCCGC 5495 1654 CCGCCACTGCCGCCGCCGCT 5496 1655 CGCCACTGCCGCCGCCGCTG 5497 1656 GCCACTGCCGCCGCCGCTGC 5498 1657 CCACTGCCGCCGCCGCTGCT 5499 1658 CACTGCCGCCGCCGCTGCTG 5500 1659 ACTGCCGCCGCCGCTGCTGC 5501 1660 CTGCCGCCGCCGCTGCTGCC 5502 1661 TGCCGCCGCCGCTGCTGCCT 5503 1662 GCCGCCGCCGCTGCTGCCTC 5504 1663 CCGCCGCCGCTGCTGCCTCC 5505 1664 CGCCGCCGCTGCTGCCTCCG 5506 1665 GCCGCCGCTGCTGCCTCCGC 5507 1666 CCGCCGCTGCTGCCTCCGCC 5508 1667 CGCCGCTGCTGCCTCCGCCG 5509 1668 GCCGCTGCTGCCTCCGCCGC 5510 1669 CCGCTGCTGCCTCCGCCGCC 5511 1670 CGCTGCTGCCTCCGCCGCCG 5512 1671 GCTGCTGCCTCCGCCGCCGC 5513 1672 CTGCTGCCTCCGCCGCCGCG 5514 1673 TGCTGCCTCCGCCGCCGCGG 5515 1674 GCTGCCTCCGCCGCCGCGGC 5516 1675 CTGCCTCCGCCGCCGCGGCC 5517 1676 CGAGCCGCCGCCACCTTCGC 5474 1677 CCGAGCCGCCGCCACCTTCG 5473 1678 GCCGAGCCGCCGCCACCTTC 5472 1679 GGCCGAGCCGCCGCCACCTT 5471 1680 TGGCCGAGCCGCCGCCACCT 5470 1681 CTGGCCGAGCCGCCGCCACC 5469 1682 ACTGGCCGAGCCGCCGCCAC 5468 1683 TACTGGCCGAGCCGCCGCCA 5467 1684 GTACTGGCCGAGCCGCCGCC 5466 1685 AGTACTGGCCGAGCCGCCGC 5465 1686 GAGTACTGGCCGAGCCGCCG 5464 1687 GGAGTACTGGCCGAGCCGCC 5463 1688 GGGAGTACTGGCCGAGCCGC 5462 1689 CGGGAGTACTGGCCGAGCCG 5461 1690 CCGGGAGTACTGGCCGAGCC 5460 1691 GCCGGGAGTACTGGCCGAGC 5459 1692 GGCCGGGAGTACTGGCCGAG 5458 1693 GGGCCGGGAGTACTGGCCGA 5457 1694 GGGGCCGGGAGTACTGGCCG 5456 1695 GGGGGCCGGGAGTACTGGCC 5455 1696 CGGGGGCCGGGAGTACTGGC 5454 1697 CGGCATAGTTCCCCGCCTTAC 2002 1698 GGCATAGTTCCCCGCCTTAC 2003 1699 GCATAGTTCCCCGCCTTACT 2004 1700 CATAGTTCCCCGCCTTACTC 2005 1701 ATAGTTCCCCGCCTTACTCT 2006 1702 TAGTTCCCCGCCTTACTCTG 2007 1703 AGTTCCCCGCCTTACTCTGC 2008 1704 GTTCCCCGCCTTACTCTGCT 2009 1705 TTCCCCGCCTTACTCTGCTC 2010 1706 TCCCCGCCTTACTCTGCTCT 2011 1707 CCCCGCCTTACTCTGCTCTA 2012 1708 CCCGCCTTACTCTGCTCTAC 2013 1709 CCGCCTTACTCTGCTCTACC 2014 1710 CGCCTTACTCTGCTCTACCT 2015 1711 ACGGCATAGTTCCCCGCCTT 2001 1712 CACGGCATAGTTCCCCGCCT 2000 1713 TCACGGCATAGTTCCCCGCC 1999 1714 GTCACGGCATAGTTCCCCGC 1998 1715 GGTCACGGCATAGTTCCCCG 1997 1716 CGGTCACGGCATAGTTCCCC 1996 1717 ACGGTCACGGCATAGTTCCC 1995 1718 CACGGTCACGGCATAGTTCC 1994 1719 ACACGGTCACGGCATAGTTC 1993 1720 CACACGGTCACGGCATAGTT 1992 1721 ACACACGGTCACGGCATAGT 1991 1722 CACACACGGTCACGGCATAG 1990 1723 TCACACACGGTCACGGCATA 1989 1724 ATCACACACGGTCACGGCAT 1988 1725 TATCACACACGGTCACGGCA 1987 1726 GTATCACACACGGTCACGGC 1986 1727 TGTATCACACACGGTCACGG 1985 1728 TTGTATCACACACGGTCACG 1984 1729 ATTGTATCACACACGGTCAC 1983 1730 CGGCCCGAGCCTCCGTGACGAGTGC 146348 1731 GGCCCGAGCCTCCGTGACGA 146349 1732 GCCCGAGCCTCCGTGACGAG 146350 1733 CCCGAGCCTCCGTGACGAGT 146351 1734 CCGAGCCTCCGTGACGAGTG 146352 1735 CGAGCCTCCGTGACGAGTGC 146353 1736 GAGCCTCCGTGACGAGTGCC 146354 1737 AGCCTCCGTGACGAGTGCCA 146355 1738 GCCTCCGTGACGAGTGCCAC 146356 1739 CCTCCGTGACGAGTGCCACC 146357 1740 CTCCGTGACGAGTGCCACCC 146358 1741 TCCGTGACGAGTGCCACCCC 146359 1742 CCGTGACGAGTGCCACCCCC 146360 1743 CGTGACGAGTGCCACCCCCT 146361 1744 GTGACGAGTGCCACCCCCTG 146362 1745 TGACGAGTGCCACCCCCTGC 146363 1746 GACGAGTGCCACCCCCTGCT 146364 1747 ACGAGTGCCACCCCCTGCTC 146365 1748 CGAGTGCCACCCCCTGCTCC 146366 1749 GCGGCCCGAGCCTCCGTGAC 146347 1750 TGCGGCCCGAGCCTCCGTGA 146346 1751 ATGCGGCCCGAGCCTCCGTG 146345 1752 TATGCGGCCCGAGCCTCCGT 146344 1753 CTATGCGGCCCGAGCCTCCG 146343 1754 CCTATGCGGCCCGAGCCTCC 146342 1755 TCCTATGCGGCCCGAGCCTC 146341 1756 CTCCTATGCGGCCCGAGCCT 146340 1757 GCTCCTATGCGGCCCGAGCC 146339 1758 GGCTCCTATGCGGCCCGAGC 146338 1759 GGGCTCCTATGCGGCCCGAG 146337 1760 TGGGCTCCTATGCGGCCCGA 146336 1761 ATGGGCTCCTATGCGGCCCG 146335 1762 CATGGGCTCCTATGCGGCCC 146334 1763 CCATGGGCTCCTATGCGGCC 146333 1764 TCCATGGGCTCCTATGCGGC 146332 1765 CTCCATGGGCTCCTATGCGG 146331 1766 CCTCCATGGGCTCCTATGCG 146330 1767 CTGGGAGGGGATCCCTCACCGAGAG 3328 1768 TGGGAGGGGATCCCTCACCG 3329 1769 GGGAGGGGATCCCTCACCGA 3330 1770 GGAGGGGATCCCTCACCGAG 3331 1771 GAGGGGATCCCTCACCGAGA 3332 1772 AGGGGATCCCTCACCGAGAG 3333 1773 GGGGATCCCTCACCGAGAGT 3334 1774 GGGATCCCTCACCGAGAGTT 3335 1775 GGATCCCTCACCGAGAGTTA 3336 1776 GATCCCTCACCGAGAGTTAG 3337 1777 ATCCCTCACCGAGAGTTAGA 3338 1778 TCCCTCACCGAGAGTTAGAA 3339 1779 CCCTCACCGAGAGTTAGAAA 3340 1780 CCTCACCGAGAGTTAGAAAA 3341 1781 CTCACCGAGAGTTAGAAAAG 3342 1782 TCACCGAGAGTTAGAAAAGC 3343 1783 CACCGAGAGTTAGAAAAGCT 3344

Hot Zones (Relative upstream location to gene start site)  650-1600 1900-2200 2900-3250 3800-4350 4800-6350 6500-7050

Examples

In FIG. 26, Both KR1 (51) and KR2 (52) demonstrated a dose-dependent inhibition response in BxPC3 (human pancreatic cancer cell line), albeit the dose response in KR1 (51) was more subtle. As would be expected, both KR1 (51) and KR2 (52) at 5 μM showed the lowest inhibition while KR1 (51) and KR2 (52) at 30 μM showed the greatest inhibition. Both KR1 (51) and KR2 (52) (FIG. 28 and FIG. 29) fit the independent and dependent DNAi motif claims.

In FIG. 27, A549 (human lung cancer cell line), KR1 shows significant (P<0.05) inhibition at 10 μM. Neither KR0525 nor the negative control demonstrates significant inhibition. Only KR1 (FIG. 28) fits the independent and dependent DNAi motif claims. KR0525's (FIG. 29) lack of inhibition is attributable to: 1) the linear base of the secondary structure either prior to or at the base of the hairpin does not contain a CG pair, 2) its secondary structure does not contain four nucleotides in its base and 3) it is located too far upstream from the KRAS transcription start site (10,265 bases upstream).

The secondary structures for KR1 and KR2 are shown in FIGS. 28 and 29. Sequence 51 (KR1) is shown in FIG. 28 and Sequence 52 (KR2) is shown in FIG. 29.

The secondary structure for KR0525 is show in FIG. 30. Sequence 53 (KR0525)—No CG in 5′ linear section of the base either prior to or in the base of the hairpin; does not contain 4 nucleotides in the base; located too far from the start site

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11955) TAATCAACAAAGCATTCATGGAGAAAATAGGTCTTATTCTAAATCTTGAA TGAGAGAGAATTGTAGCAAACAGAAAGACAAAAAGGTGCTGGGTGAGAAA AGGAGCAGAGACATAAATAAAATATCCAATTTTAAGGGTATAGAGAGGGG ATTCACTCAAGGAGGGGAGACCATCTATCTGCTTTGAGAAGCTGGGAAAC AAAGTCATAGGGTCAGGATGGTGCCTGACTATGGATGCTCTCAAAAGCTA GGCACCAAGGATTTGGACTGGATTCAGCTGGATATAAGAAGTTATTACAG ACTTGGAAGCAAGATTAAGTCTCCGGGAAGGGGAGACTTAACTGGGACCA GAGATCATTTTCCCCTATAATTTTAAAGGTACTTATCATCTTTAGGTACT CATTAGGTACTTAGCTTGTAACTCTTTCCACTGTTCAAATATATACCCAG TATGCATGTAGCCTATATGGAGCAGGCACAGAGTAAATGTTTGATGATGA TAAAAGACATGCGGAAGAAAGGTTAATTTGGCAACATCATAAAACTGAAT TGAGACAAAGAAAGCCAGGAGGTAGGAAAGTCAATGAAGAAGTTATTCCA GAAATGTAGCTGAGAAGGAAGGAATACAGAAGAGGCAGATATGGGAAAAT ACTCAGGAAGTATAATTAAAAGGAGCTGTGACTAATTTTAATAAGGACTG GGTTAAAAATTAAGTTTTCATGTCTAAATGTTCTGGAGGACCATGATGTC ACTCAGGTAAGATGGAGGAATTGAGAGAGGGAATTCGTTAGAGGGAATAA CATGGGGAATTTGGCTTTGGACAGGCATTTGCCATGATAACAGAATATTC ATTTAGAAATGGTCCAGGAAATTGGTTTGATGAGAATGAAGTGCCTGTGA AGAGAGAGGACTGAAGCTTGTTATAATTTCATTCACTTCAGGAATATTTA CAGAGGACCCAAATGTGCTAAGAACTATGAAAACATAGAATTAAAAGAAA TGGGCCTGGAATAATTTACAACCTAGTAAAGCAGTTATGGGAAAACATAT TTGCAAAAAAGGTATACAAAGTATAATGAAATAAGTGTCCAGTAAGGATA AAGTGCAGAGTAAGTGAATTAAGCAGCACCCATTCATGTGTTCAAATTCC TGCCAGAGTCAAAAGGTTGTGCTGAAGTAGAGTCCATGAAAGCATCGTAG ATGGCTCCTCCTGCTCAAGTTCCCCTGCTCTGCGTCCTGCTACTCTGGCC ACAACCGTCTGGACCCAGGGTTGACACACAAACAAACACAATAATCTTTT AGCCAGACATAAAGAAGGCCAGCCACCAATCAGGAAAATTGTGTCCCATA AAGGCCCTTCCTATTGAACAGTGAATGACAGACATGGCCAGATCTTCTCT CTTGGAATGCTTTGAATGTTAGTCACAGAGAGTGACCACTAGAAGCACAG ATAGCAGTAGAAGCTAAGACTACATGAAAAAGCAGTGGACAGATGGTGAT TTATGAGAATGGCAAAATTACTAGAGTCATAGGCAATGGATACTTGTTAA TGAAGGGATGAGCAGGGCCCCACAGCCTGTTGCTGGCTCACAAGTGCAGT TGATTGCTGGACTGAACAGCAGCTCTCCGCCTGATGATAGGGTTTTTTAA AGTGTCCTTATTGCCTTAAAGTAAATCCTCAGCATTTGCAGTGCTCTGAG GGTGTCCTAGCATTTTATACCTTTTTTCTAAGAGCCCAGGTAACATAAGG GTACTCCTGTTGTTCTGGCTTTAATTCTATCTGCAGAAGAGGGTTTCTTG TGAAAGAAAGGGTCAGTATGGTCTTTTATCTGTACAGCAGATAAAAAGGG TATGTACGTGCACACCTTTGTACGTGGCTGCCTTCCCAGGACAGTCTGAC AGTAGAGGGTAGAAACTTCAGTTGTAGCTGAGAGCAGGCCTGGAATCCCC ATGCTTATACTTTTTATTTCCTCCCCCCTTTCCCATTGTGATCACAGGCT ACTTCAGTGTGCTTGTCCTTGGAGAGAGCAAGGGAAGGGAGAGCCAGGGA GACTGTTCAAGGGAGCCACCAGGCTCGAGAAAGAGGAACCCCTGAAGACA GTAGAAAGTGCAGGTGCCAAGAATTTGAATATCTACATCAGAGTTTCTCA ATGTGCACACAGTGAACTACCAGTTTAGGATCATTTGATTTGCTAAAAAT GAAGATTACTGGTCTACCTTAGACCAACTGAATAAAATATCTGGGTGAGG GGCCTAGGAACTTGCATTTTTGGTAGGCATGGCAGGTGATTCCTAAAGCA TTTACCCTTGAGACCTCTATGTTAAGGAAAGAAAGGTAATGTTGCAAGGA GGTGGTGCCGGCTTCTAAGAAAGTACCCAGGACTGAACGGCAGAAAGACC TGACATACCATATGTATAAATTGCTGTGGAAGTGAAAAGGAAAGAGAAAG TGTCTGAGGTAAAACTGGAGTGTGGGGTGCGTGGAACAAATGGTTGGATG CAGATTTGCTTTACGAATCATGAGCCTAGATGATAACTGAGACCATGTGG ATGGATTAGGTTTCTGCTAATGCCAGAATTTTTATAATCAGCATAAAAGT GCTATATAAAGCTTTCCCCTCTTCTATATTATAGTCCTTTTAAGATGTAT GGAACATCAACTATAGGAAGAACATCATATTCACAGCTGTAAGAGGAAAC AAGAACTTATCATGCACTTGATGTTGTACAAAATAAATCTGTGATTTATG CTTGAGTGACCACAAAGTAGCATACACATAAGCGCAAATTCATTCATTTA AGAATTCCTTGTGTCTATTATGTACGAGATAAGTATCTCTGAGCTGCACG GAATGTGGCTTATCAGAAGGTGACCTAAGTTTCAAAGCAGATTTTGTTAA GATGAAGACAGAGATTGACAGGAGGTTTAAGACACTCTGTCTAAAGTAAA GATTTAGAGTCACAGAGTTCATGGATTAGGATTTAGAATCCACAGAGGGT CCACAGATTCACTCATTCAACATTCCATAAATATTTATTGAATGCCTTTT TGTGTCAGAGACTGTCTTAGGTGCTGGAAATTTAGCAGTAAATGAAACAG ACCAAAACCCATGCCCTCATGGAGCTTACATTCTGATGGTAGAGAGACAA GAAAACAAAATAGATAGTGTATTATTGAAGGTGATGAGAGCTCTGGAGAA AAAGTAGGAAAAGAGACAGATCTGGGACAAGGGCGAAATTACAGTATCAA AGATGATCTTTTTAGGGAAGATCTCCTTTTAAAAACACTTTGGAACAAAG ATTTAAATGAGGTGCCAGAGGGGTAGCAAGTGCATATTCCCTGAGGAAGA CGCCTGCCTGGCATTTTCAAGGAACAGCCAGTAACCAATGTTTATCTACG TAAGTAAGGAAGGGAGAACAGTAGGATGAGAGTTCAGAGAAGAGGGTAGG GGATATCAAATAATTTAAGGCCATGTAGGATTTTTGAGAAGAATTTTGCT TTTATGTCAAGTGGAATGAGGGCCACTGATGATCTGGGAGTAGAGTGACT ATGATCCGACATGAAGTATACTCCATTTTTTAACTATGTGAACTTGTGCC AACGTTTTAACCTCTAAATCTGTTTCGTCATTTGTAAAACGGTAAAAAGT ATTTTACCTCATAAGGTTGTCGTGATGATTAAATAAGATGATACGATAAG TGCAAAAGATTTAGCTTGTACTTAACATAGAGTAGGCACATTTTCTCCCC TTCCCTGTCTTTCACTTTTCTCTTCTGCCCCTTCCACCTGGCGCTAGGAG GGGGAGACTGGAATAAACCTTGCAGATTACAGCCCGTGTAAGAGTAGAAA GGAAAGGATGACAGTTGATGTAAAGCCTTGGTTAACAGACATAATAGCTG GGATTTAAATTCAGCTTTATTGGTGGTTTATGATGTGGACTAGAGGAATG GAACTGAAAGTCTCGGAGGAGGGGCGATCCTATCAGGTACAGGCGCTGCT TTTCCAGCCCTCAATCCTCAAGACTCTCCCAAGATACATTTCTAGGTAGT TTATCAACACAGACTCCGGGTATGCTAGCATGTTTAATTGCCCCATTGTT TAATGTCTTAACTCCACGAACTTTAACTGATTAATCTGTCTTCTAATTAA TGTTTGAATGACTCTCCTCAGGTCTAAACTACCAAGGCCATCTCTACTTA AAAACAGTTGTCTTTTGTTTGTGATTTCAGGGGCCCTGGGTATAAGCGAA GTCCCTGTTTAGAGACCTTGTGATGGGTTCAAAATATCAAGAAAGATAGC AAAATATCACAAGCCTCCTGACCCGAGAAGATTAGCGTTGAAAGGGTCTG TCGTGTTTGTTTGGGCCTGGGGCTAAATTCCCAGCCCAAGTGCTGAGGCT GATAATAATCGGGGCGGCGATCAGACAGCCCCGGTGTGGGAAATCGTCCG CCCGGTCTCCCTAAGTCCCCGAAGTCGCCTCCCACTTTTGGTGACTGCTT GTTTATTTACATGCAGTCAATGATAGTAAATGGATGCGCGCCAGTATAGG CCGACCCTGAGGGTGGCGGGGTGCTCTTCGCAGCTTCTCTGTGGAGACCG GTCAGCGGGGCGGCGTGGCCGCTCGCGGCGTCTCCCTGGTGGCATCCGCA CAGCCCGCCGCGGTCCGGTCCCGCTCCGGGTCAGAATTGGCGGCTGCGGG GACAGCCTTGCGGCTAGGCAGGGGGCGGGCCGCCGCGTGGGTCCGGCAGT CCCTCCTCCCGCCAAGGCGCCGCCCAGACCCGCTCTCCAGCCGGCCCGGC TCGCCACCCTAGACCGCCCCAGCCACCCCTTCCTCCGCCGGCCCGGCCCC CGCTCCTCCCCCGCCGGCCCGGCCCGGCCCCCTCCTTCTCCCCGCCGGCG CTCGCTGCCTCCCCCTCTTCCCTCTTCCCACACCGCCCTCAGCCGCTCCC TCTCGTACGCCCGTCTGAAGAAGAATCGAGCGCGGAACGCATCGATAGCT CTGCCCTCTGCGGCCGCCCGGCCCCGAACTCATCGGTGTGCTCGGAGCTC GATTTTCCTAGGCGGCGGCCGCGGCGGCGGAGGCAGCAGCGGCGGCGGCA GTGGCGGCGGCGAAGGTGGCGGCGGCTCGGCCAGTACTCCCGGCCCCCGC CATTTCGGACTGGGAGCGAGCGCGGCGCAGGCACTGAAGGCGGCGGCGGG GCCAGAGGCTCAGCGGCTCCCAGGTGCGGGAGAGAGGTACGGAGCGGACC ACCCCTCCTGGGCCCCTGCCCGGGTCCCGACCCTCTTTGCCGGCGCCGGG CGGGGCCGGCGGCGAGTGAATGAATTAGGGGTCCCCGGAGGGGCGGGTGG GGGGCGCGGGCGCGGGGTCGGGGCGGGCTGGGTGAGAGGGGTCTGCAGGG GGGAGGCGCGCGGACGCGGCGGCGCGGGGAGTGAGGAATGGGCGGTGCGG GGCTGAGGAGGGTGAGGCTGGAGGCGGTCGCCGCTGGTGCTGCTTCCTGG ACGGGGAACCCCTTCCTTCCTCCTCCCCGAGAGCCGCGGCTGGAGGCTTC TGGGGAGAAACTCGGGCCGGGCCGGCTGCCCCTCGGAGCGGTGGGGTGCG GTGGAGGTTACTCCCGCGGCGCCCCGGCCTCCCCTCCCCCTCTCCCCGCT CCCGCACCTCTTGCCTCCCTTTCCAGCACTCGGCTGCCTCGGTCCAGCCT TCCCTGCTGCATTTGGCATCTCTAGGACGAAGGTATAAACTTCTCCCTCG AGCGCAGGCTGGACGGATAGTGGTCCTTTTCCGTGTGTAGGGGATGTGTG AGTAAGAGGGGAGGTCACGTTTTGGAAGAGCATAGGAAAGTGCTTAGAGA CCACTGTTTGAGGTTATTGTGTTTGGAAAAAAATGCATCTGCCTCCGAGT TCCTGAATGCTCCCCTCCCCCATGTATGGGCTGTGACATTGCTGTGGCCA CAAAGGAGGAGGTGGAGGTAGAGATGGTGGAAGAACAGGTGGCCAACACC CTACACGTAGAGCCTGTGACCTACAGTGAAAAGGAAAAAGTTAATCCCAG ATGGTCTGTTTTGCTTGGTCAAGTTAAACCCGAAGAAAACCCGCAGAGCA GAAGCAAGGCTTTTTCCTTGCTAGTTGAGTGTAGACAGCAATAGCAAAAA TAGTACTTGAAGTTTAATTTACCTGTTCTTGTCCTTTCCCCTATTTCTTA TGTATTACCCTCATCCCCTCGTCTCTTTTATACTACCCTCATTTTGCAGA TGTGTTCTACATCTCAAGAGTTATTACAGTACTCCAAAACAGCACTTACA TGATTTTTTAAACTTACAGAGGAATTGTAGCAATCCACCAGCTAACCGCC TGAAATAGACTTAAACATGTGCATCTCCTTTTTTTTTTTTTTTTTGAGAC ACAGTCTCGCTCTGTTGCCCAGGCTGGAGTGCAATGGCGCGGTATCGGCT CACTGAAACCTCCGCCTCCTGGGTTCAAGCAATTCTCCTGCCTCAGCCTC CCGAGTAGCTGGGACTAGTAGGTGCACGCCACCATGCCCAGCTAATTTTT GTATTTTTAGTAGAGACAGAGTTTCATCATGTTGGTCAGGATGGTCTCCA TCTGCTCTGTTGCCCAGGCTGGAGTGCAGTGGCGCCGTCTCGGCTCACTG CAACCTCTGCCTCCTGCATTCAAGCAATTCTCCTGCCTCAGCCTCCCGAA TAACTGGGATTACAGGTGTCTGCTGCCATGCCCGGCTAATTTTTTGTATT TTTAGTAGAGACGGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAACTC CTGACCTCGTGATCTGCCCGCCTCGGCCTCCCACAGTGGCATGTGCATCT TATAGCTGAAGTCTAAGCCTTCTTAAATCTTGAGATCCATCAAAACAGAC AGGTTTTCTAATTGTTATACAATGTATATGTTATGTTTATAATAGAAATC ATTTTACAAATAAGTTATAAATGGGAAAGGTCTATTTGTAATTATCAGCT CAGAATTAACCATAAAACTGGTGTCACTGAAGTGACTGAGGTCCAAAATG CTGACTCTGCATGTTATAGACTACAGATATCAAATATGGTTGCTAACAAT AGTTTACTTTGAGACTGTAGCCATCCACAGTATATTTGCTTTTAAGAGAT GGTAGATGGTAATTCAGTTTTATGAAAAATAAAAATGAATTTTCTTCCAT TACAAAATTGTTGGATTCGAGTCCAGTCCACTCCTTACTAGCTTTTCTAA CTCTCGGTGAGGGATCCCCTCCCAGCCCATGATCTTCATTTGGTAAGACT CCTTTGGAACCCAGTTCTCTCTAGTGGATTTAAATGTGATTTGGTTTTAA AAATCTCATTCAAGGAATTTTTTTTTTTTCTGGAAACAACCACCGCATAA ACAAGTAAACCGGAAGATACATGTGGCTCTGAATTCATATATATACACAA ACTCTAATCCAATGTCTGTCCACAGTATTTCCTAGGCTAGTAAACTTTTT GGCCTTAACGACCCCTCTACCCTCTTTGTTTTTTTGAGAGAGAGAGTCTC ACTCTGTCACCCAGGCCGGAATGCAGTGGCGCGATCTCGGCCCGCTACTA CCTCCGACTCTCAGGCTCAAGCGATTCTCCCGCCTCAGCTTCCCGAGTAG CCGGGATTACAGGCTCCCGCCACCGGGCTAATTGTATTTTTAGATACGGG ATTTCACCATGTTGGCCAGGCTGGTCTCGACCTCCTGACCTCAGGTGATC CGCCCGCCTAAGCCTCCCAAAGTGCTGGGATTACAGGCCACCACACCCGG CCTACACTCTTAAAAATTATCGAAGGGGCCGGGCACATTGGCTCTTATCT GTAATCCCAGCACTTTGGGAGACTGAGGCGGGAGGATCGCTTGAGGCCAG GAGTTGGAGACCAGCGTACTCAACATAGTGAGACCTTGTTATAAAGAAAA AAAAAATCCAGGATTAAAAAAAATCTTTGATTTGTTTGGGATTTATTAAT ATTTACCGTATTGGAAATTAAAACAATTTTTTAAAATGTATTCATTTAAA AATAATAAGCCCATTACTTGGTAACATGAATAAAATATTTTATGAAAAAT AACTATTTTCCAAAACAAAACCAAAACTTAGAAAAGTGGTATTGTTTCAC ACTTCAGTAAATCTCTTTAATGATGTGGCTTAATAGAAGATATGGATTCT TATATCTGCATCTGCATTCAATCTATTATGATCACACATCTGGAAAACTT GTGAAAGAATGGGAGTTAAAAGGGTAAAGGACATCTTAATGTTATTATGA AAACAGTTTTGACCTCTTGCACACCAGAAAAGTCTTAGTAACCTGAGGGG TTCCTAGACCACATTTTGAGAACTGTTTTAGGCTATGCAAACTGGTTGGG GGGAGGTTGGGGTAGGCAGAGAGCTAGAAGATACATTTTAGTGTAATTCT CCTCATCTATTCCTAATTGCTTTGGCCTACATTTGAAATAAAGCGTGGAG GCAAACGGGATAAGATACATGTTTGTAGTGGTTGTTAACTTCACCCTAGA CAAGCAGCCAATAAGTCTAGGTAGAGCAGAGTAAGGCGGGGAACTATGCC GTGACCGTGTGTGATACAATTTTTCTAGCCTGTGGTGCTTTTTGCGGCAG GGCTTAGGAGTAAGGTTAGTATGTTATCATTTGGGAAACCAAATTATTAT TTTGGGTCTTCAGTCAATTATGATGCTGTGTATATTTAGTGTTTATCTAC AATATATGCACATTCATTAATTTGGAGCTACTCATCCTATAATAAATAGT TGTGCATTTACTCCCATTTTTTTCTGCATTTCTCTCCTTATTTATAATTA TGTGTTACATGAGGGAAAGGAGGTGAAATTAAACATTCATATTATTTCAA AAAATTTGAAACAACTAACTAAAAAATATGTTTTATTTTCTGTATGGTGT TTGTTATACAATCTGTCAATATTCATGCACCTCTTGGGAGACAGTGTATG AAAAGCAAAGAGTAACAGTCACATGGATTACTGATTACTGAGATATATTC ACTTGCATCTTTTTTTTTTTTTGAGACGGAGTGGCTCTGTCGCCCAGGCT GGAGTGCAGTGGCGTGATCTCGGCTCACTGCAAGCTCCGCCTCCTGGGTT CACGCCATTCTTCTGCCTCAGCCTCCCAAGTAGCTGGGACTACAGGCGCC CGCCACCACGCCCGGCTAATTTTTTTATATTTTTAGTAGAGACGGGGTTT CACCGGGTTAGCCAGGATGGTCTTGATCTCCTGACCTCGTGATCCACCCT CCTCGGCCTCCCAAAGTGCTAGGATTATAGGCGTGAGCCACCGTGCCCGG CTCACTTGCATCTCTTAACAGCTGTTTTCTTACTAAAAACAGTGTTTATC TCTAATCTTTTTGTTTGTTTGTTTGTTTTGAGATGGAGTCTTACTCCGTC ACCCAATCTGGAGTGCAGTGGCGTGATCTGGGCTCACTGCAACCTCTGCC TCCCGGGTTCAAGTGATTCTCCTTCCTCAGCCTCCCCAGTAGCTAGGACT ACAGGAGAGCGCCACCACGCCTGATTAATTTTTGTATTTTTAGTAGAGAG AGGGTTTCACCATATTGGCCAGGCTGGTCTTGAACTCCTGGCCTCAGGTG ATCCACCCGCCTTGGCCTCTGAAAGTGCTGGGATTACAGGCATGAGCCGC CGCACCCGGCTTTCTAATCTTTATCTTTTTTTGTGCAGCGGTGATACAGG ATTATGTATTGTACTGAACAGTTAATTCGGAGTTCTCTTGGTTTTTAGCT TTATTTTCCCCAGAGATTTTTTTTTTTTTTTTTTTTTTTGAGACGGAGTC TTGCTCTATCGCCAGGCTGGAGTGCAGTGGCGCCATCTCGGCTCATTGCA ACCTCGGACTCCTATTTTCCCCAGAGATATTTCACACATTAAAATGTCGT CAAATATTGTTCTTCTTTGCCTCAGTGTTTAAATTTTTATTTCCCCATGA CACAATCCAGCTTTATTTGACACTCATTCTCTCAACTCTCATCTGATTCT TACTGTTAATATTTATCCAAGAGAACTACTGCCATGATGCTTTAAAAGTT TTTCTGTAGCTGTTGCATATTGACTTCTAACACTTAGAGGTGGGGGTCCA CTAGGAAAACTGTAACAATAAGAGTGGAGATAGCTGTCAGCAACTTTTGT GAGGGTGTGCTACAGGGTGTAGAGCACTGTGAAGTCTCTACATGAGTGAA GTCATGATATGATCCTTTGAGAGCCTTTAGCCGCCGCAGAACAGCAGTCT GGCTATTTAGATAGAACAACTTGATTTTAAGATAAAAGAACTGTCTATGT AGCATTTATGCATTTTTCTTAAGCGTCGATGGAGGAGTTTGTAAATGAAG TACAGTTCATTACGATACACGTCTGCAGTCAACTGGAATTTTCATGATTG AATTTTGTAAGGTATTTTGAAATAATTTTTCATATAAAGGTGAGTTTGTA TTAAAAGGTACTGGTGGAGTATTTGATAGTGTATTAACCTTATGTGTGAC ATGTTCTAATATAGTCACATTTTCATTATTTTTATTATAAGGCCTGCTGA AA ATG

7) MTTP. Microsomal triglyceride transfer protein is an an essential chaperone for the biosynthesis/lipoprotein assembly of apolipoprotein B (apoB)-containing triglyceride-rich lipoproteins Inhibition of MTTP prevents the assembly of apo B-containing lipoproteins by inhibiting chylomicrons and VLDL synthesis. As a result, decreases in plasma levels of LDL-C are observed (Shoulders et al., Hum Mol Genet 2 (12): 2109-16). Patients carry mutations in the MTTP gene exhibit abetalipoproteinemia resulting from the loss of its lipid transfer activity.

MTTP is also recognized to play a role in the biosynthesis of CD1, glycolipid presenting molecules, as well as in the regulation of cholesterol ester biosynthesis. Recently, MTTP has been implicated in the propagation of hepatitis C virus, where the virus hijacks lipoprotein assembly for its secretion. Therefore, MTTP is a good target to lower plasma lipids and treat disorders characterized by higher production of apoB-containing lipoproteins such as atherosclerosis, metabolic syndrome, familial combined hyperlipidemia, homozygous and heterozygous familial hypercholesterolemia and hypertriglyceridemia (reviewed in Hussain et al. Nutrition & Metabolism 2012, 9:14). MTTP is also recognized to be involved in the immune response against foreign lipid antigens, such that targeting it may also be useful for modulating the inflammatory response during T cell mediated processes such as inflammatory bowel disease, autoimmune hepatitis and asthma (Hussain et al., Curr Opin Lipidol 2008, 19:277-284). Current therapies that inhibit MTTP without increasing hepatic lipids and plasma transaminases are lacking.

Protein: MTTP Gene: MTTP (Homo sapiens, chromosome 4, 100485240-100545154 [NCBI Reference Sequence: NC_(—)000004.11]; start site location: 100496067; strand: positive)

Gene Identification GeneID 4547 HGNC 7467 HPRD 01144 MIM 157147

Targeted Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 1784 AACCGCCGTAGCCTCCACTGCG 10855 1870 TGGCCGCAGTTCGATGACGTAAGACG 10828

Target Shift Sequences Relative upstream Sequence location to gene ID No: Sequence (5′-3′) start site 1784 AACCGCCGTAGCCTCCACTGCG 10855 1785 ACCGCCGTAGCCTCCACTGC 10856 1786 CCGCCGTAGCCTCCACTGCG 10857 1787 CGCCGTAGCCTCCACTGCGT 10858 1788 GCCGTAGCCTCCACTGCGTA 10859 1789 CCGTAGCCTCCACTGCGTAA 10860 1790 CGTAGCCTCCACTGCGTAAC 10861 1791 GTAGCCTCCACTGCGTAACT 10862 1792 TAGCCTCCACTGCGTAACTA 10863 1793 AGCCTCCACTGCGTAACTAC 10864 1794 GCCTCCACTGCGTAACTACC 10865 1795 CCTCCACTGCGTAACTACCG 10866 1796 CTCCACTGCGTAACTACCGC 10867 1797 TCCACTGCGTAACTACCGCC 10868 1798 CCACTGCGTAACTACCGCCC 10869 1799 CACTGCGTAACTACCGCCCC 10870 1800 ACTGCGTAACTACCGCCCCT 10871 1801 CTGCGTAACTACCGCCCCTG 10872 1802 TGCGTAACTACCGCCCCTGC 10873 1803 GCGTAACTACCGCCCCTGCC 10874 1804 CGTAACTACCGCCCCTGCCT 10875 1805 GTAACTACCGCCCCTGCCTC 10876 1806 TAACTACCGCCCCTGCCTCT 10877 1807 AACTACCGCCCCTGCCTCTG 10878 1808 ACTACCGCCCCTGCCTCTGG 10879 1809 CTACCGCCCCTGCCTCTGGG 10880 1810 TACCGCCCCTGCCTCTGGGA 10881 1811 ACCGCCCCTGCCTCTGGGAA 10882 1812 CCGCCCCTGCCTCTGGGAAT 10883 1813 CGCCCCTGCCTCTGGGAATT 10884 1814 CAACCGCCGTAGCCTCCACT 10854 1815 GCAACCGCCGTAGCCTCCAC 10853 1816 CGCAACCGCCGTAGCCTCCA 10852 1817 ACGCAACCGCCGTAGCCTCC 10851 1818 GACGCAACCGCCGTAGCCTC 10850 1819 AGACGCAACCGCCGTAGCCT 10849 1820 AAGACGCAACCGCCGTAGCC 10848 1821 TAAGACGCAACCGCCGTAGC 10847 1822 GTAAGACGCAACCGCCGTAG 10846 1823 CGTAAGACGCAACCGCCGTA 10845 1824 ACGTAAGACGCAACCGCCGT 10844 1825 GACGTAAGACGCAACCGCCG 10843 1826 TGACGTAAGACGCAACCGCC 10842 1827 ATGACGTAAGACGCAACCGC 10841 1828 GATGACGTAAGACGCAACCG 10840 1829 CGATGACGTAAGACGCAACC 10839 1830 TCGATGACGTAAGACGCAAC 10838 1831 TTCGATGACGTAAGACGCAA 10837 1832 GTTCGATGACGTAAGACGCA 10836 1833 AGTTCGATGACGTAAGACGC 10835 1834 CAGTTCGATGACGTAAGACG 10834 1835 GCAGTTCGATGACGTAAGAC 10833 1836 CGCAGTTCGATGACGTAAGA 10832 1837 CCGCAGTTCGATGACGTAAG 10831 1838 GCCGCAGTTCGATGACGTAA 10830 1839 GGCCGCAGTTCGATGACGTA 10829 1840 TGGCCGCAGTTCGATGACGT 10828 1841 ATGGCCGCAGTTCGATGACG 10827 1842 AATGGCCGCAGTTCGATGAC 10826 1843 AAATGGCCGCAGTTCGATGA 10825 1844 GAAATGGCCGCAGTTCGATG 10824 1845 CGAAATGGCCGCAGTTCGAT 10823 1846 TCGAAATGGCCGCAGTTCGA 10822 1847 TTCGAAATGGCCGCAGTTCG 10821 1848 GTTCGAAATGGCCGCAGTTC 10820 1849 GGTTCGAAATGGCCGCAGTT 10819 1850 GGGTTCGAAATGGCCGCAGT 10818 1851 CGGGTTCGAAATGGCCGCAG 10817 1852 GCGGGTTCGAAATGGCCGCA 10816 1853 TGCGGGTTCGAAATGGCCGC 10815 1854 TTGCGGGTTCGAAATGGCCG 10814 1855 ATTGCGGGTTCGAAATGGCC 10813 1856 CATTGCGGGTTCGAAATGGC 10812 1857 CCATTGCGGGTTCGAAATGG 10811 1858 TCCATTGCGGGTTCGAAATG 10810 1859 TTCCATTGCGGGTTCGAAAT 10809 1860 CTTCCATTGCGGGTTCGAAA 10808 1861 TCTTCCATTGCGGGTTCGAA 10807 1862 TTCTTCCATTGCGGGTTCGA 10806 1863 TTTCTTCCATTGCGGGTTCG 10805 1864 CTTTCTTCCATTGCGGGTTC 10804 1865 CCTTTCTTCCATTGCGGGTT 10803 1866 CCCTTTCTTCCATTGCGGGT 10802 1867 CCCCTTTCTTCCATTGCGGG 10801 1868 TCCCCTTTCTTCCATTGCGG 10800 1869 CTCCCCTTTCTTCCATTGCG 10799 1870 TGGCCGCAGTTCGATGACGTAAGACG 10828 1871 GGCCGCAGTTCGATGACGTA 10829 1872 GCCGCAGTTCGATGACGTAA 10830 1873 CCGCAGTTCGATGACGTAAG 10831 1874 CGCAGTTCGATGACGTAAGA 10832 1875 GCAGTTCGATGACGTAAGAC 10833 1876 CAGTTCGATGACGTAAGACG 10834 1877 AGTTCGATGACGTAAGACGC 10835 1878 GTTCGATGACGTAAGACGCA 10836 1879 TTCGATGACGTAAGACGCAA 10837 1880 TCGATGACGTAAGACGCAAC 10838 1881 CGATGACGTAAGACGCAACC 10839 1882 GATGACGTAAGACGCAACCG 10840 1883 ATGACGTAAGACGCAACCGC 10841 1884 TGACGTAAGACGCAACCGCC 10842 1885 GACGTAAGACGCAACCGCCG 10843 1886 ACGTAAGACGCAACCGCCGT 10844 1887 CGTAAGACGCAACCGCCGTA 10845 1888 GTAAGACGCAACCGCCGTAG 10846 1889 TAAGACGCAACCGCCGTAGC 10847 1890 AAGACGCAACCGCCGTAGCC 10848 1891 AGACGCAACCGCCGTAGCCT 10849 1892 GACGCAACCGCCGTAGCCTC 10850 1893 ACGCAACCGCCGTAGCCTCC 10851 1894 CGCAACCGCCGTAGCCTCCA 10852 1895 GCAACCGCCGTAGCCTCCAC 10853 1896 CAACCGCCGTAGCCTCCACT 10854 1897 AACCGCCGTAGCCTCCACTG 10855 1898 ACCGCCGTAGCCTCCACTGC 10856 1899 CCGCCGTAGCCTCCACTGCG 10857 1900 CGCCGTAGCCTCCACTGCGT 10858 1901 GCCGTAGCCTCCACTGCGTA 10859 1902 CCGTAGCCTCCACTGCGTAA 10860 1903 CGTAGCCTCCACTGCGTAAC 10861 1904 GTAGCCTCCACTGCGTAACT 10862 1905 TAGCCTCCACTGCGTAACTA 10863 1906 AGCCTCCACTGCGTAACTAC 10864 1907 GCCTCCACTGCGTAACTACC 10865 1908 CCTCCACTGCGTAACTACCG 10866 1909 CTCCACTGCGTAACTACCGC 10867 1910 TCCACTGCGTAACTACCGCC 10868 1911 CCACTGCGTAACTACCGCCC 10869 1912 CACTGCGTAACTACCGCCCC 10870 1913 ACTGCGTAACTACCGCCCCT 10871 1914 CTGCGTAACTACCGCCCCTG 10872 1915 TGCGTAACTACCGCCCCTGC 10873 1916 GCGTAACTACCGCCCCTGCC 10874 1917 CGTAACTACCGCCCCTGCCT 10875 1918 GTAACTACCGCCCCTGCCTC 10876 1919 TAACTACCGCCCCTGCCTCT 10877 1920 AACTACCGCCCCTGCCTCTG 10878 1921 ACTACCGCCCCTGCCTCTGG 10879 1922 CTACCGCCCCTGCCTCTGGG 10880 1923 TACCGCCCCTGCCTCTGGGA 10881 1924 ACCGCCCCTGCCTCTGGGAA 10882 1925 CCGCCCCTGCCTCTGGGAAT 10883 1926 CGCCCCTGCCTCTGGGAATT 10884 1927 ATGGCCGCAGTTCGATGACG 10827 1928 AATGGCCGCAGTTCGATGAC 10826 1929 AAATGGCCGCAGTTCGATGA 10825 1930 GAAATGGCCGCAGTTCGATG 10824 1931 CGAAATGGCCGCAGTTCGAT 10823 1932 TCGAAATGGCCGCAGTTCGA 10822 1933 TTCGAAATGGCCGCAGTTCG 10821 1934 GTTCGAAATGGCCGCAGTTC 10820 1935 GGTTCGAAATGGCCGCAGTT 10819 1936 GGGTTCGAAATGGCCGCAGT 10818 1937 CGGGTTCGAAATGGCCGCAG 10817 1938 GCGGGTTCGAAATGGCCGCA 10816 1939 TGCGGGTTCGAAATGGCCGC 10815 1940 TTGCGGGTTCGAAATGGCCG 10814 1941 ATTGCGGGTTCGAAATGGCC 10813 1942 CATTGCGGGTTCGAAATGGC 10812 1943 CCATTGCGGGTTCGAAATGG 10811 1944 TCCATTGCGGGTTCGAAATG 10810 1945 TTCCATTGCGGGTTCGAAAT 10809 1946 CTTCCATTGCGGGTTCGAAA 10808 1947 TCTTCCATTGCGGGTTCGAA 10807 1948 TTCTTCCATTGCGGGTTCGA 10806 1949 TTTCTTCCATTGCGGGTTCG 10805 1950 CTTTCTTCCATTGCGGGTTC 10804 1951 CCTTTCTTCCATTGCGGGTT 10803 1952 CCCTTTCTTCCATTGCGGGT 10802 1953 CCCCTTTCTTCCATTGCGGG 10801 1954 TCCCCTTTCTTCCATTGCGG 10800 1955 CTCCCCTTTCTTCCATTGCG 10799

Hot Zones (Relative upstream location to gene start site) 10750-10900

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11956) TCTTGAAAATAATCTGTCCTCTCTATCTAGTTCCTTTAAATATCTTCTCT CTCTCTCTGATATTCTGCAGTTTAATTATGATGTATCTACTTGTGTGTAT GTGTGTTTTAAAAATTATCCTGCTTAAGACTTATTGAGCCTCGTGAATCT GTGGATTGGTATCTGTGATAGGCAGACAATGGCTCCCCAAAGATCTTAAA TGTCTTAATCTCCAGAACCTGTGATAGTCTAAGTTAAGGTTGTAGATGAA ATTAAAGTTACCAATCCACAGACCTCAGGGTAAAAAGATTATCCTGGATT ATTTAGGCAGGCCCAGTATAATCACAAGGATTCATAAAACGGAAGAGGGA GACAGAAGAGATGGTCAGAGTGATATGAAGTAAAAAGGATTCAGCTTACT CTTGCTGGATTTGTAAATGCAGGAAGGGACCACGAGTCAAGGAATGCAGG TAGCTTCTAGAAGCTAGAAAAGGTAAGAAACAAATTCTCCCCTAAAGCCT CCAGAAAGGGATACACCTGCCAATACTTTCATTTTATCCCTGTGAGACCA GTGTTAGACTTCTGACCTCCAAGAGTATAAGACAATAAATCTGCTGTTTT AAGCCACTAAGTTTTGTGGTAATTTGTTATGGTAGCTATGGAGAACTGAT ACAGTGCCTTTCAATAGTTCTTGGAAATTCTTCAAATATATTCCCCAAAT ATTGCCTTTGCACCACTCACTCTATCCTCTATATCTCTTGACCTCTCTTT AACATTTTTTATTTTCTTATTTTGGTAATTATTTAAAACAATTGGCTTCT TGTTCCACTTCAAATAAATTCATATTTTTATATCTACATATTAAGAATTA GTTCATAAATGTAATTGTTGTATCGTATATACTTTAAAAGGAAAATTGCA TTTATACTTGGATATTATTATATTTTAGGTTTTGAAATTTCTTTTTTTAA ATGTCTGATAAATTTATTCTGATCAAAATTAAAATCTTATTGTATTTACA GTAGTCTACTAAAAGAGTTTACATCAATTCTCTCCTTTAAATGCTAGCAA TTCAGTTTTGTGTGGTCAAAGATAATTTAGAATCCTTTTATGGTAACTGA TATGATACAAAGGATTTTTTTGGTTGTTGTCAAATTTGTATGTGCGTATA TATGTATAGGGGGTAGAAAATTTGGTTAGTGACTCTATTTTGGAAAAATG AATGTTCTTTTTGGAGTTTTAGATTCCCAGTGTTTCAAACCAAGTTTGCT TTTGATAAGGAATTCAGTGAATCTTTATTTTCTCGTAGAGAATTTTTAAA CAATACCTTTCAAAATATTGTATGTATTCCTATATAATTTTGCCTTGGAA TAAAAAAAGTCATCACATTAAGAATATTTTTAACTTAAGAAATTTTTTCA AGATTCTTTCAAAATGCTAACTCCCTCATTGATTTGAAAATACTTTTAGA GTTTAAACTATTATGACCATGTAGGGATAACTTAAAGTGTCCTTCTAAAT TTTTTTTTCATTAATGTCATGCTTCTTTCTAAGAACAAAGTGTTTCAATG TTATAAACAGACTGTTTTTTCACATAGCTTTCATACATCCTGACTTTCTA TATCTAGTAGGGTAATACATTTCCTTCCAACCTTTAGTGGGGTGAAGTTC ATTTGTCTTCTAATCTAAGAAAATACGTTAACTCCTGACAACCTCTGACA CTCAACAGAAACACAACTGTACTTTGGAATTAATCATCCATCTTTATTTC ATAGTCTCCTTATTTATCAATGCAAATGGAAGTATAGCAAATATTTCACA GTGTCATGTACTACCTAAGGAATTTCATTGCAATAGCATTGTTTTGAAAT GTATCATTTGATTTTGATCTAACTATCATAGATCAGTTTTACCCCTGTAT GTTCAGCCTGAATGTCTAACCACAATTTCACAAAAATCAAGGGCTCTGTT AGTTTTCATACAAAATTGTGATGACTTATTATGAAGACAGTCCCAATTAA CAATTGCATGTCATCTGTAAGAAATCAATTTTTTTCTCACTCCCCACCTG TAATTTTTTTTAAGCAAAGAAGAATACTTGGGTCTAGGTTTCAAGGTTTA TTTTTCTGTAGTCCTAATATCACTTCAGTAAATTAATCTGAGACTCATTT TTCTAATATGTCAAATGACTGTTGTAAGGATTATTATAAAAACGTAAGGT GATTAACAAAGTATGTAATTGTTCCATAGATGGCAGCTCTTGTTTTTTTA CTGTGCTTTGCTTCTGCCCTGCTTCACATATTATGTAAAATAGCTGGTGA GTTTTAGGAGGTAGTGCTCACATGTTTGCACAGTGTTTGCTGTGGACGAT CAACAGTAACAGAAGAGCATACTTCTTCGATACACAGAGTTTACTGAATT TGGAAAGGCTTTGGCATTCTTATGTCATCTGTAATGAAACAATCTCCAGA AGTCTTTTCTAAAATGTCCTTGTAAAAAGAAAAAAGTTATGTTTATATTT TATAAAAGGATGATGTTATTTATAACCAGCAGAAGCAGCCTTATTTGAAC ATCTTATGTTGAAATTGCTACTTAATACAGTGACTCATAGGAGCTTTCTA GTGGAAATCAAATGCTCAAATGAAATAGAATTTAGTTTGTTAGGCAATAG TGATATGTCTTTTATTGGTTGGACTCTGGAAAACACTTGACAACGAATAG TACTTTACCCGAAGGGCACGTATCATGCACCACATAGCCTAACCACAAAC ATTAAAGGTCTTGTAACTGTGAGCCTCAAATGAAAATACATAAGGACAGC TCTCATACAATCAAATACAATACAAACTAGCTTTTAATTTAAATAAATAT GTAAGTAAAGTTCAAGTGACTATAATGATTTTATATCCCTATGTATGTAT CACAGAAATTGTGGCAAACTGTAGAAATCTATTCAAATGGAAAGTAACAA AGCACTTTCACATTGCCTTGTATTCAAAATCCCTACTCTTCATAAAAACT TATATTTCTTTAACAAAGCTACTTTTCTGTTTAACTCCCGGAAAACTTCG TATTTATAACTTAAGGGGGTTTCTCCAACCAAACAATTTATTTTTGCTAG GTACTATAGCTATATTTTTTATACAAAATTTGTGACAGCAAATGAAATTC TAATCCCAATAGAAGAACAAACAATTTTCATGTTTCGATCTTCATATATA TAATTCAAGAGGAAATATGCTTAACTTTGTAGATTTTTACATTTTAAATT GCATTGTGTCTGTATCAAGTCTACTATCTTTTACCTAGATTGTCTGGAAG ATTTAAGCTCAAGGTTACGGTTTGAGAAAAGGGTTTTGAGAGTGACCAGG ATAGATTTAAGAATTCATTTTATACTAAAATATGGCCATAAATATTTTTA AATACATTCAAATAGCCCTTTGCTGGCACATTTTTTCCCTTCTTTGCCAA AACATTCCCACAGGCGGCCTAAGTCACCTCATTTTATAGGTTTAGTAGGT TTAGCAGGCTTTATGTGCTCTAGTAGGGTTAGTAGGTTTTGTTCATATCA GGTCTCTCTCATGGGAGTTTCCAGGGACAAGGATTGCTTCAGTTAGTATG GCCTTAGCCATACTAGGGTATTTGCTTTAATTCTACAGAAGTTTTCTAAT TAATATTCTGTAGCAAAAGAACTAAGATCTGGAATTCCCCCTCTTAATCT CTTCCTAGAAATGAGATTCAGAAAGGACAGGACTGCATCCAGCCTGTTTG GGAACTCAGACAAATGTGTGTTGTCACAGACACAAATAGAGGTCTACTAT GAAATAATTGGCTTGCTAGTGTGCTAATGACAGACAATGCTGATTTGCTC CAACCTCATACAGTTTCACACATAAGGACAATCATCTATGTTTCATGAAA GTTCTATCTACTTTAACATTATTTTGAAGTGATTGGTGGTGGTATGAATT AACAGTTTAAATTTAAATCCTAAAATTCAGTGTGAATTTTTTATAATAGC ATAAAAATTCAAAGATGTCCATACAAGAAAAATTAAAATTTGGTTAGGTT TAGCAGAGTTTGAGAATCCTTACTACCCTCCCACATAGTATTGTAATGTG AATATAGGCAGTTACTATTACAGGCATAATGATGATTATGTATTAAGCAG AAAGAAGTATCACCACCAGTTTTTTTCTTTGAATGCCCCTCAGTACTTCT GCATTTATAGGATGGTAGACTGGTTTGGTTTAGCTCTCAAAAGTGAAAAC ATTTAAAGTTTCCTCATTGGGTGAAAAAAATTAAAAAGAGTGAGAGACTG AAAACTGCAGCCCACCTACGTTTAATCATTAATAGTGAGCCCTTCAGTGA ACTTAGGTCCTGATTTTGGAGTTTGGAGTCTGACCTTTCCCCAAAGATAA ACATGATTGTTGCAGGTTCTGAAGAGGGTCACTCCCTCACTGGCTGCCAT TGAAAGAGTCCACTTCTCAGTGACTCCTAGCTGGGCACTGGATGCAGTTG AGGATTGCTGGTCAAT ATG

8) ApoC III. Apolipoprotein C-III is a protein component of very low density lipoprotein (VLDL). APOC3 inhibits lipoprotein lipase and hepatic lipase; it is thought to inhibit hepatic uptake of triglyceride-rich particles (reviewed in Mendevil et al., Arteriosclerosis, Thrombosis and Vascular Biology 30 (2): 239-45). The APOA1, APOC3 and APOA4 genes are closely linked in both rat and human genomes. The A-I and A-IV genes are transcribed from the same strand, while the A-1 and C-III genes are convergently transcribed. An increase in apoC-III levels induces the development of hypertriglyceridemia. Two novel susceptibility haplotypes (specifically, P2-S2-X1 and P1-S2-X1) have been discovered in ApoAI-CIII-AIV gene cluster on chromosome 11q23; these confer approximately threefold higher risk of coronary heart disease in normal as well as non-insulin diabetes mellitus. Apo-CIII delays the catabolism of triglyceride rich particles. Elevations of Apo-CIII found in genetic variation studies may predispose patients to non-alcoholic fatty liver disease.

ISIS-APOCIIIRx is an antisense drug designed to reduce apolipoprotein C-III, or apoC-III, protein production and lower triglycerides. ApoC-III regulates triglyceride metabolism in the blood and is an independent cardiovascular risk factor. People who do not produce apoC-III have lower levels of triglycerides and lower instances of cardiovascular disease. ApoC-III is elevated in patients with dyslipidemia, or an abnormal concentration of lipids in the blood, and is frequently associated with multiple metabolic abnormalities, such as insulin resistance and/or metabolic syndrome. In human population studies, lower levels of apoC-III and triglycerides correlated with a lower rate of cardiovascular events. In certain populations, apoC-III mediates insulin resistance, which can make metabolic syndrome worse.

Protein: ApoC-III Gene: APOC3 (Homo sapiens, chromosome 11, 116700624-116703787 [NCBI Reference Sequence: NC_(—)000011.9]; start site location: 116701299; strand: positive)

Gene Identification GeneID 345 HGNC 610 HPRD 00132 MIM 107720

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 1956 GAGTCGGTGGTCCAGGAGGGGCCGC 1614 1957 CTGCGGCTGAGGTGTCATTCGTGACTCAG 4214 1992 GCGGGCGGGTGAGACAGAAGCGCC 4130 1993 CCTCGCGAGCGTGGGTGCACGC 3985 2028 CGATGTCTCCCTCGAGATCACA 3717 2054 GGACGGACGGATATCTGAGGCCAG 2195 2062 CGTCCCCGCCACGTTGAAAGGC 3954 2089 TCTCGGACATGCTCAAATGGTGCAGGCG 4080 2108 CACCGACAGGAGCCAATAGTGCAACG 4065 2127 GTCCGGCAGAGGGACCCATGCTGACG 4940 2136 CGTGAGGCACATGTCCGTGTG 3511 2170 CAGATGCAGCAAGCGGGCGGGAGAG 798 2176 CCACGCTGCTGTCCCGCCAGCCCTGCAG 848 2206 ACCCGCCCCCACCCTGTGTGCCCCCACCC 1276 GCCCCCACCCTGTGTGCCCCC 2225 CGCTCAGAGCCCGAGGCCTTTG 1352

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 1956 GAGTCGGTGGTCCAGGAGGGGCCGC 1614 1957 CTGCGGCTGAGGTGTCATTCGTGACTCAG 4214 1958 TGCGGCTGAGGTGTCATTCG 4215 1959 GCGGCTGAGGTGTCATTCGT 4216 1960 CGGCTGAGGTGTCATTCGTG 4217 1961 GGCTGAGGTGTCATTCGTGA 4218 1962 GCTGAGGTGTCATTCGTGAC 4219 1963 CTGAGGTGTCATTCGTGACT 4220 1964 TGAGGTGTCATTCGTGACTC 4221 1965 GAGGTGTCATTCGTGACTCA 4222 1966 AGGTGTCATTCGTGACTCAG 4223 1967 GGTGTCATTCGTGACTCAGT 4224 1968 GTGTCATTCGTGACTCAGTC 4225 1969 TGTCATTCGTGACTCAGTCT 4226 1970 GTCATTCGTGACTCAGTCTC 4227 1971 TCATTCGTGACTCAGTCTCC 4228 1972 CATTCGTGACTCAGTCTCCT 4229 1973 ATTCGTGACTCAGTCTCCTC 4230 1974 TTCGTGACTCAGTCTCCTCC 4231 1975 TCGTGACTCAGTCTCCTCCT 4232 1976 CGTGACTCAGTCTCCTCCTC 4233 1977 ACTGCGGCTGAGGTGTCATT 4213 1978 AACTGCGGCTGAGGTGTCAT 4212 1979 AAACTGCGGCTGAGGTGTCA 4211 1980 CAAACTGCGGCTGAGGTGTC 4210 1981 TCAAACTGCGGCTGAGGTGT 4209 1982 GTCAAACTGCGGCTGAGGTG 4208 1983 GGTCAAACTGCGGCTGAGGT 4207 1984 AGGTCAAACTGCGGCTGAGG 4206 1985 GAGGTCAAACTGCGGCTGAG 4205 1986 GGAGGTCAAACTGCGGCTGA 4204 1987 TGGAGGTCAAACTGCGGCTG 4203 1988 CTGGAGGTCAAACTGCGGCT 4202 1989 CCTGGAGGTCAAACTGCGGC 4201 1990 TCCTGGAGGTCAAACTGCGG 4200 1991 GTCCTGGAGGTCAAACTGCG 4199 1992 GCGGGCGGGTGAGACAGAAGCGCC 4130 1993 CCTCGCGAGCGTGGGTGCACGC 3985 1994 CTCGCGAGCGTGGGTGCACG 3986 1995 TCGCGAGCGTGGGTGCACGC 3987 1996 CGCGAGCGTGGGTGCACGCA 3988 1997 GCGAGCGTGGGTGCACGCAT 3989 1998 CGAGCGTGGGTGCACGCATG 3990 1999 GAGCGTGGGTGCACGCATGG 3991 2000 AGCGTGGGTGCACGCATGGG 3992 2001 GCGTGGGTGCACGCATGGGC 3993 2002 CGTGGGTGCACGCATGGGCT 3994 2003 GTGGGTGCACGCATGGGCTG 3995 2004 TGGGTGCACGCATGGGCTGT 3996 2005 GGGTGCACGCATGGGCTGTG 3997 2006 GGTGCACGCATGGGCTGTGC 3998 2007 GTGCACGCATGGGCTGTGCC 3999 2008 TGCACGCATGGGCTGTGCCA 4000 2009 GCACGCATGGGCTGTGCCAG 4001 2010 CACGCATGGGCTGTGCCAGT 4002 2011 ACGCATGGGCTGTGCCAGTC 4003 2012 CGCATGGGCTGTGCCAGTCC 4004 2013 CCCTCGCGAGCGTGGGTGCA 3984 2014 CCCCTCGCGAGCGTGGGTGC 3983 2015 TCCCCTCGCGAGCGTGGGTG 3982 2016 GTCCCCTCGCGAGCGTGGGT 3981 2017 GGTCCCCTCGCGAGCGTGGG 3980 2018 AGGTCCCCTCGCGAGCGTGG 3979 2019 CAGGTCCCCTCGCGAGCGTG 3978 2020 GCAGGTCCCCTCGCGAGCGT 3977 2021 AGCAGGTCCCCTCGCGAGCG 3976 2022 CAGCAGGTCCCCTCGCGAGC 3975 2023 GCAGCAGGTCCCCTCGCGAG 3974 2024 GGCAGCAGGTCCCCTCGCGA 3973 2025 AGGCAGCAGGTCCCCTCGCG 3972 2026 AAGGCAGCAGGTCCCCTCGC 3971 2027 AAAGGCAGCAGGTCCCCTCG 3970 2028 CGATGTCTCCCTCGAGATCACA 3717 2029 GATGTCTCCCTCGAGATCAC 3718 2030 ATGTCTCCCTCGAGATCACA 3719 2031 TGTCTCCCTCGAGATCACAC 3720 2032 GTCTCCCTCGAGATCACACA 3721 2033 TCTCCCTCGAGATCACACAG 3722 2034 CTCCCTCGAGATCACACAGG 3723 2035 TCCCTCGAGATCACACAGGC 3724 2036 CCCTCGAGATCACACAGGCC 3725 2037 CCTCGAGATCACACAGGCCT 3726 2038 CTCGAGATCACACAGGCCTT 3727 2039 TCGAGATCACACAGGCCTTT 3728 2040 CGAGATCACACAGGCCTTTC 3729 2041 GCGATGTCTCCCTCGAGATC 3716 2042 GGCGATGTCTCCCTCGAGAT 3715 2043 AGGCGATGTCTCCCTCGAGA 3714 2044 GAGGCGATGTCTCCCTCGAG 3713 2045 AGAGGCGATGTCTCCCTCGA 3712 2046 GAGAGGCGATGTCTCCCTCG 3711 2047 GGAGAGGCGATGTCTCCCTC 3710 2048 TGGAGAGGCGATGTCTCCCT 3709 2049 TTGGAGAGGCGATGTCTCCC 3708 2050 CTTGGAGAGGCGATGTCTCC 3707 2051 GCTTGGAGAGGCGATGTCTC 3706 2052 GGCTTGGAGAGGCGATGTCT 3705 2053 AGGCTTGGAGAGGCGATGTC 3704 2054 GGACGGACGGATATCTGAGGCCAG 2195 2055 GACGGACGGATATCTGAGGC 2196 2056 ACGGACGGATATCTGAGGCC 2197 2057 CGGACGGATATCTGAGGCCA 2198 2058 GGACGGATATCTGAGGCCAG 2199 2059 GACGGATATCTGAGGCCAGG 2200 2060 ACGGATATCTGAGGCCAGGA 2201 2061 CGGATATCTGAGGCCAGGAG 2202 2062 CGTCCCCGCCACGTTGAAAGGC 3954 2063 GTCCCCGCCACGTTGAAAGG 3955 2064 TCCCCGCCACGTTGAAAGGC 3956 2065 CCCCGCCACGTTGAAAGGCA 3957 2066 CCCGCCACGTTGAAAGGCAG 3958 2067 CCGCCACGTTGAAAGGCAGC 3959 2068 CGCCACGTTGAAAGGCAGCA 3960 2069 GCCACGTTGAAAGGCAGCAG 3961 2070 CCACGTTGAAAGGCAGCAGG 3962 2071 CACGTTGAAAGGCAGCAGGT 3963 2072 ACGTTGAAAGGCAGCAGGTC 3964 2073 CGTTGAAAGGCAGCAGGTCC 3965 2074 ACGTCCCCGCCACGTTGAAA 3953 2075 CACGTCCCCGCCACGTTGAA 3952 2076 TCACGTCCCCGCCACGTTGA 3951 2077 GTCACGTCCCCGCCACGTTG 3950 2078 GGTCACGTCCCCGCCACGTT 3949 2079 AGGTCACGTCCCCGCCACGT 3948 2080 CAGGTCACGTCCCCGCCACG 3947 2081 ACAGGTCACGTCCCCGCCAC 3946 2082 AACAGGTCACGTCCCCGCCA 3945 2083 TAACAGGTCACGTCCCCGCC 3944 2084 TTAACAGGTCACGTCCCCGC 3943 2085 ATTAACAGGTCACGTCCCCG 3942 2086 CATTAACAGGTCACGTCCCC 3941 2087 TCATTAACAGGTCACGTCCC 3940 2088 TTCATTAACAGGTCACGTCC 3939 2089 TCTCGGACATGCTCAAATGGTGCAGGCG 4080 2090 CTCGGACATGCTCAAATGGT 4081 2091 TCGGACATGCTCAAATGGTG 4082 2092 CGGACATGCTCAAATGGTGC 4083 2093 CTCTCGGACATGCTCAAATG 4079 2094 GCTCTCGGACATGCTCAAAT 4078 2095 TGCTCTCGGACATGCTCAAA 4077 2096 ATGCTCTCGGACATGCTCAA 4076 2097 GATGCTCTCGGACATGCTCA 4075 2098 GGATGCTCTCGGACATGCTC 4074 2099 TGGATGCTCTCGGACATGCT 4073 2100 GTGGATGCTCTCGGACATGC 4072 2101 GGTGGATGCTCTCGGACATG 4071 2102 TGGTGGATGCTCTCGGACAT 4070 2103 CTGGTGGATGCTCTCGGACA 4069 2104 TCTGGTGGATGCTCTCGGAC 4068 2105 CTCTGGTGGATGCTCTCGGA 4067 2106 ACTCTGGTGGATGCTCTCGG 4066 2107 CACTCTGGTGGATGCTCTCG 4065 2108 CACCGACAGGAGCCAATAGTGCAACG 4876 2109 ACCGACAGGAGCCAATAGTG 4877 2110 CCGACAGGAGCCAATAGTGC 4878 2111 CGACAGGAGCCAATAGTGCA 4879 2112 TCACCGACAGGAGCCAATAG 4875 2113 CTCACCGACAGGAGCCAATA 4874 2114 ACTCACCGACAGGAGCCAAT 4873 2115 CACTCACCGACAGGAGCCAA 4872 2116 GCACTCACCGACAGGAGCCA 4871 2117 TGCACTCACCGACAGGAGCC 4870 2118 CTGCACTCACCGACAGGAGC 4869 2119 ACTGCACTCACCGACAGGAG 4868 2120 CACTGCACTCACCGACAGGA 4867 2121 GCACTGCACTCACCGACAGG 4866 2122 GGCACTGCACTCACCGACAG 4865 2123 AGGCACTGCACTCACCGACA 4864 2124 CAGGCACTGCACTCACCGAC 4863 2125 TCAGGCACTGCACTCACCGA 4862 2126 GTCAGGCACTGCACTCACCG 4861 2127 GTCCGGCAGAGGGACCCATGCTGACG 4940 2128 TCCGGCAGAGGGACCCATGC 4941 2129 CCGGCAGAGGGACCCATGCT 4942 2130 CGGCAGAGGGACCCATGCTG 4943 2131 GGTCCGGCAGAGGGACCCAT 4939 2132 TGGTCCGGCAGAGGGACCCA 4938 2133 GTGGTCCGGCAGAGGGACCC 4937 2134 TGTGGTCCGGCAGAGGGACC 4936 2135 GTGTGGTCCGGCAGAGGGAC 4935 2136 CGTGAGGCACATGTCCGTGTG 3511 2137 GTGAGGCACATGTCCGTGTG 3512 2138 TGAGGCACATGTCCGTGTGA 3513 2139 GAGGCACATGTCCGTGTGAC 3514 2140 AGGCACATGTCCGTGTGACC 3515 2141 GGCACATGTCCGTGTGACCT 3516 2142 GCACATGTCCGTGTGACCTG 3517 2143 CACATGTCCGTGTGACCTGC 3518 2144 ACATGTCCGTGTGACCTGCC 3519 2145 CATGTCCGTGTGACCTGCCT 3520 2146 ATGTCCGTGTGACCTGCCTG 3521 2147 TGTCCGTGTGACCTGCCTGT 3522 2148 GTCCGTGTGACCTGCCTGTC 3523 2149 TCCGTGTGACCTGCCTGTCC 3524 2150 CCGTGTGACCTGCCTGTCCC 3525 2151 CGTGTGACCTGCCTGTCCCT 3526 2152 ACGTGAGGCACATGTCCGTG 3510 2153 TACGTGAGGCACATGTCCGT 3509 2154 ATACGTGAGGCACATGTCCG 3508 2155 CATACGTGAGGCACATGTCC 3507 2156 GCATACGTGAGGCACATGTC 3506 2157 AGCATACGTGAGGCACATGT 3505 2158 AAGCATACGTGAGGCACATG 3504 2159 GAAGCATACGTGAGGCACAT 3503 2160 TGAAGCATACGTGAGGCACA 3502 2161 TTGAAGCATACGTGAGGCAC 3501 2162 CTTGAAGCATACGTGAGGCA 3500 2163 CCTTGAAGCATACGTGAGGC 3499 2164 CCCTTGAAGCATACGTGAGG 3498 2165 CCCCTTGAAGCATACGTGAG 3497 2166 GCCCCTTGAAGCATACGTGA 3496 2167 GGCCCCTTGAAGCATACGTG 3495 2168 GGGCCCCTTGAAGCATACGT 3494 2169 AGGGCCCCTTGAAGCATACG 3493 2170 CAGATGCAGCAAGCGGGCGGGAGAG 798 2171 CCAGATGCAGCAAGCGGGCG 797 2172 TCCAGATGCAGCAAGCGGGC 796 2173 GTCCAGATGCAGCAAGCGGG 795 2174 TGTCCAGATGCAGCAAGCGG 794 2175 GTGTCCAGATGCAGCAAGCG 793 2176 CCACGCTGCTGTCCCGCCAGCCCTGCAG 848 2177 CACGCTGCTGTCCCGCCAGC 849 2178 ACGCTGCTGTCCCGCCAGCC 850 2179 CGCTGCTGTCCCGCCAGCCC 851 2180 GCTGCTGTCCCGCCAGCCCT 852 2181 CTGCTGTCCCGCCAGCCCTG 853 2182 TGCTGTCCCGCCAGCCCTGC 854 2183 GCTGTCCCGCCAGCCCTGCA 855 2184 CTGTCCCGCCAGCCCTGCAG 856 2185 TGTCCCGCCAGCCCTGCAGC 857 2186 GTCCCGCCAGCCCTGCAGCC 858 2187 TCCCGCCAGCCCTGCAGCCC 859 2188 CCCGCCAGCCCTGCAGCCCA 860 2189 CCGCCAGCCCTGCAGCCCAG 861 2190 CGCCAGCCCTGCAGCCCAGA 862 2191 TCCACGCTGCTGTCCCGCCA 847 2192 GTCCACGCTGCTGTCCCGCC 846 2193 AGTCCACGCTGCTGTCCCGC 845 2194 GAGTCCACGCTGCTGTCCCG 844 2195 TGAGTCCACGCTGCTGTCCC 843 2196 CTGAGTCCACGCTGCTGTCC 842 2197 ACTGAGTCCACGCTGCTGTC 841 2198 GACTGAGTCCACGCTGCTGT 840 2199 AGACTGAGTCCACGCTGCTG 839 2200 GAGACTGAGTCCACGCTGCT 838 2201 GGAGACTGAGTCCACGCTGC 837 2202 AGGAGACTGAGTCCACGCTG 836 2203 TAGGAGACTGAGTCCACGCT 835 2204 CTAGGAGACTGAGTCCACGC 834 2205 CCTAGGAGACTGAGTCCACG 833 2206 ACCCGCCCCCACCCTGTGTGCCCCCCG 1276 2207 CCCGCCCCCACCCTGTGTGC 1277 2208 CCGCCCCCACCCTGTGTGCC 1278 2209 CGCCCCCACCCTGTGTGCCC 1279 2210 CACCCGCCCCCACCCTGTGT 1275 2211 CCACCCGCCCCCACCCTGTG 1274 2212 CCCACCCGCCCCCACCCTGT 1273 2213 CCCCACCCGCCCCCACCCTG 1272 2214 CCCCCACCCGCCCCCACCCT 1271 2215 CCCCCCACCCGCCCCCACCC 1270 2216 GCCCCCCACCCGCCCCCACC 1269 2217 AGCCCCCCACCCGCCCCCAC 1268 2218 CAGCCCCCCACCCGCCCCCA 1267 2219 GCAGCCCCCCACCCGCCCCC 1266 2220 AGCAGCCCCCCACCCGCCCC 1265 2221 CAGCAGCCCCCCACCCGCCC 1264 2222 CCAGCAGCCCCCCACCCGCC 1263 2223 CCCAGCAGCCCCCCACCCGC 1262 2224 ACCCAGCAGCCCCCCACCCG 1261 2225 CGCTCAGAGCCCGAGGCCTTTG 1352 2226 GCTCAGAGCCCGAGGCCTTT 1353 2227 CTCAGAGCCCGAGGCCTTTG 1354 2228 TCAGAGCCCGAGGCCTTTGC 1355 2229 CAGAGCCCGAGGCCTTTGCC 1356 2230 AGAGCCCGAGGCCTTTGCCC 1357 2231 GAGCCCGAGGCCTTTGCCCC 1358 2232 AGCCCGAGGCCTTTGCCCCT 1359 2233 GCCCGAGGCCTTTGCCCCTC 1360 2234 CCCGAGGCCTTTGCCCCTCC 1361 2235 CCGAGGCCTTTGCCCCTCCC 1362 2236 CGAGGCCTTTGCCCCTCCCT 1363 2237 CCGCTCAGAGCCCGAGGCCT 1351 2238 GCCGCTCAGAGCCCGAGGCC 1350 2239 GGCCGCTCAGAGCCCGAGGC 1349 2240 AGGCCGCTCAGAGCCCGAGG 1348 2241 AAGGCCGCTCAGAGCCCGAG 1347 2242 CAAGGCCGCTCAGAGCCCGA 1346 2243 CCAAGGCCGCTCAGAGCCCG 1345 2244 GCCAAGGCCGCTCAGAGCCC 1344 2245 GGCCAAGGCCGCTCAGAGCC 1343 2246 GGGCCAAGGCCGCTCAGAGC 1342 2247 AGGGCCAAGGCCGCTCAGAG 1341 2248 AAGGGCCAAGGCCGCTCAGA 1340 2249 GAAGGGCCAAGGCCGCTCAG 1339 2250 AGAAGGGCCAAGGCCGCTCA 1338 2251 GAGAAGGGCCAAGGCCGCTC 1337

Hot Zones (Relative upstream location to gene start site) 700-900 1100-1400 1550-1700 2100-2300 3450-4300 4700-5000

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11957) GAGACATAACCATTGTACCTGCCTCCTAGGCTGTGAGGATTCACTGAGAT GATCTTATAGTGCTTGCAACAATGTCTGGCACATAGGAAAAGTGATCACT AAATGTTAGCCACGTCTTACTCCTGCAAGGCTCACCTCCCTGGAACCCAT CGGTCCCAACCCTGCTCCTGAATCAGGCACAGTCCAGCTTGCAGCGGGAG CAAAGGTCAGTACTCAGTGCCCCTGTCCCTTCCCCAGGCCAGAGGGGAGG AGGAGACTGAGTCACGAATGACACCTCAGCCGCAGTTTGACCTCCAGGAC TTACAGTCCTAGCAGCCGGTGCCACTAGCATGTGAGAGGTCCAGAGGCGC TTCTGTCTCACCCGCCCGCCTGGGTGCACCCATGCTGGGAGCGCCTGCAC CATTTGAGCATGTCCGAGAGCATCCACCAGAGTGTGTGTGGATTCACAGA AGTGTGCAAATCACTAAGAACCAAGGGACTGGCACAGCCCATGCGTGCAC CCACGCTCGCGAGGGGACCTGCTGCCTTTCAACGTGGCGGGGACGTGACC TGTTAATGAATGTATTTACTTCCCAAAGTCTGAGGGTACGTTTTGCATCA ATCTGTAGATGGATTTGTTTTGGGGAGCAGGGAGAGAATGAGAGCCCCCT GTGCTCAGTCTTAGAGGGTGCAAGTAGCTGATGGGAAGAGCAGACTGCCT TCCAGCCAGGCCTGGTCCTGTGAGTCAGGGACGTCCATCTTAGTGGGCAT GAAAGGCCTGTGTGATCTCGAGGGAGACATCGCCTCTCCAAGCCTCTCCT TATCTGTGCAACAGGCAGACTTAATGATTGGTGAGGCAATGAGGCTGATA GCTCAGCATTAGCTACAGCCACCCCTCCTGGCCAACCACACAGGGATCAA ACCAGGGGTCAGTCCAGAGGTCAGAGTCAGGAGCAGACAACTCAGATCCA GCCAGGGACAGGCAGGTCACACGGACATGTGCCTCACGTATGCTTCAAGG GGCCCTCCCCCGGGCAGAACTGAAGGACAGCTCCTGTTGCCATAGGAGGG AGCTGGGTGAGATACTAGGAGGAACTTCCGGCATGATGATGTGTGATGAA CAAGGGCCTCTGGCCAACAGGTCTGAATCAGGGCTGCCCAGCCCAGCCTG GTGGGAAGGGCATGGAGCATGGGGGCTCATGTACTAAACCTCACCTGGAC ACAAGGTGAAACAGCCCAACCCCAGAGGACCATTTTTGGCCCCGGATGGT CAAATCCCCTCTTCCTCCCATCTACCACTGGCTTCTCCCTGGAGCAGTCT TCATCCCAGGGGAGCCATGATGGGAGAGAGGGGCAGCGCAGGCTGGCCAC CAAGAGATCCCCTGCCGGGGTGCAGGTTGGACTGTTGGTGAGGGGCCACA GGTATTCTCAGGTACCAAGCCCTTGGAAGGAGACAAGGTACCAGGCTTCC TGGAGGTGTGCTACATCTAGCTCAGCACCCTGCCAGGTCTCTCTACCCAC ATGTCCTGACCTCCCTGGGTCCGTTGCCATGCGGGAGAGAGAGGCCAGGC TCCTCCAGACCCTCTGCAGAGATGGAAAGGCTTGGAGGGTCTGGGGCCAC GGGACCCCGCCAGCCCATTCTAGCACACCCGGGCCCATAGACCTTGTTGC CTGCCCCTGCCTGGATCTGGGTCCCCACTGTGCCTTTGCCTCTGGGGCTA TGGAGCAGGCCGCAGCAGAAGAGGAAAGGGCATCCCCAATACCAAATCCT CCAGTGACCACTTCTTCACCTTCTACCCCACCACCAAAGTCTGCAGGAGA CTTGAGACAGGTTTGTTCTGGGCGTGTGACTGATGCCTCTATAGGGGTCT CAGTGCTCTAAGCCGTCTGGTATTTGCCTGGGGTGTGTGAAGACCTGGAT TAAGGTTCCCAGCCTTACTACTAATGGGCTGTGCACTTGGAGCCCTTAGA GCCTTAGGTTTCTAACCTATAAAATGGACTTAACGTCTACTTCACAGGGT TCTATTTGCATTTTAACAGAAAACAAAGTCTTAAGTCAAAGGAATGAATC TCTCTCTCTCTCTCTCTCTCTCTTTTTTAGACCAAGTCTAGCTCTGTCAC TGGAGTGCAATGGTGCGATCTCTGCTCACTGCAACCTCCACCTCCGGGGT TCAAGCAATTCTCGTGCCTCAGCCTCCTGAGTAGCTGGGACTACAGGCGT GCATCACCATGCTCGGCTAATTTTTTGTATTTTTAGTAGAGACTGGGTTT CGCCATGTTGCCCAGGCTGGTCTCGAACTCCTGGCCTCAGATATCCGTCC GTCCCAGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCACTGTGCCCA GCCAGGAATGGATCGCTAATAGAGGAATTCCAAGTCTCACCCACCGATAA AGAATTCTGAGGGCAGAGCCGGGCCACTTTCTCAGGCCTCTGATTTCATA CTGTGGTGTTAGTTACTTCTGAGAGGACAGCTTGCTGCCAGAGCTCTATT TTTTATGTTAGAGGCTCCTTCTGCCTGCAGACTCTGCTGTCTGGGAAGGG CACAGCGTTAGGAGGGAGAGGGAGGTGTGAGTCCCTCCATGGACCCGCTG CTTTGTACTTCTCTATCTCATTTCCTTTTCAGCACCACTCTGGGCAATCA GTATTCCAGCCCCATTTTATCCTCAGAAAATTGAGGCTCTGAGATGTTAT CTCTGTGACCTGGGTCCTATTACGTGCCAAAGGCATCATTTAAGCCTAAG ATGTCCTGGCTCCAAGGTGTCAGCATCTGGAAGACAGGCGCCCTCATCCT GCCATCCCTGCTGCGGCTTCACTGTGGGCCCAGGGGACATCTCAGCCCCG AGAAGGGTCAGCGGCCCCTCCTGGACCACCGACTCCCCGCAGAACTCCTC TGTGCCCTCTCCTCACCAGACCTTGTTCCTCCCAGTTGCTCCCACAGCCA GGGGGCAGTGAGGGCTGCTCTTCCCCCAGCCCCACTGAGGAACCCAGGAA GGTGAACGAGAGAATCAGTCCTGGTGGGGGCTGGGGAGGGCCCCAGACAT GAGACCAGCTCCTCCCCCAGGGGATGTTATCAGTGGGTCCAGAGGGCAAA ATAGGGAGCCTGGTGGAGGGAGGGGCAAAGGCCTCGGGCTCTGAGCGGCC TTGGCCCTTCTCCACCAACCCCTGCCCTACACTAAGGGGGAGGCAGCGGG GGGCACACAGGGTGGGGGCGGGTGGGGGGCTGCTGGGTGAGCAGCACTCG CCTGCCTGGATTGAAACCCAGAGATGGAGGTGCTGGGAGGGGCTGTGAGA GCTCAGCCCTGTAACCAGGCCTTGCCGGAGCCACTGATGCCTGGTCTTCT GTGCCTTTACTCCAAACACCCCCCAGCCCAAGCCACCCACTTGTTCTCAA GTCTGAAGAAGCCCCTCACCCCTCTACTCCAGGCTGTGTTCAGGGCTTGG GGCTGGTGGAGGGAGGGGCCTGAAATTCCAGTGTGAAAGGCTGAGATGGG CCCGAGGCCCCTGGCCTATGTCCAAGCCATTTCCCCTCTCACCAGCCTCT CCCTGGGGAGCCAGTCAGCTAGGAAGGAATGAGGGCTCCCCAGGCCCACC CCCAGTTCCTGAGCTCATCTGGGCTGCAGGGCTGGCGGGACAGCAGCGTG GACTCAGTCTCCTAGGGATTTCCCAACTCTCCCGCCCGCTTGCTGCATCT GGACACCCTGCCTCAGGCCCTCATCTCCACTGGTCAGCAGGTGACCTTTG CCCAGCGCCCTGGGTCCTCAGTGCCTGCTGCCCTGGAGATGATATAAAAC AGGTCAGAACCCTCCTGCCTGTCTGCTCAGTTCATCCCTAGAGGCAGCTG CTCCAGGTAATGCCCTCTGGGGAGGGGAAAGAGGAGGGGAGGAGGATGAA GAGGGGCAAGAGGAGCTCCCTGCCCAGCCCAGCCAGCAAGCCTGGAGAAG CACTTGCTAGAGCTAAGGAAGCCTCGGAGCTGGACGGGTGCCCCCCACCC CTCATCATAACCTGAAGAACATGGAGGCCCGGGAGGGGTGTCACTTGCCC AAAGCTACACAGGGGGTGGGGCTGGAAGTGGCTCCAAGTGCAGGTTCCCC CCTCATTCTTCAGGCTTAGGGCTGGAGGAAGCCTTAGACAGCCCAGTCCT ACCCCAGACAGGGAAACTGAGGCCTGGAGAGGGCCAGAAATCACCCAAAG ACACACAGCATGTTGGCTGGACTGGACGGAGATCAGTCCAGACCGCAGGT GCCTTGATGTTCAGTCTGGTGGGTTTTCTGCTCCATCCCACCCACCTCCC TTTGGGCCTCGATCCCTCGCCCCTCACCAGTCCCCCTTCTGAGAGCCCGT ATTAGCAGGGAGCCGGCCCCTACTCCTTCTGGCAGACCCAGCTAAGGTTC TACCTTAGGGGCCACGCCACCTCCCCAGGGAGGGGTCCAGAGGCATGGGG ACCTGGGGTGCCCCTCACAGGACACTTCCTTGCAGGAACAGAGGTGCC AT G

9) APO B. Apolipoprotein B (ApoB) are the primary apolipoproteins of chylomicrons and low-density lipoproteins (LDL) and is required for lipoprotein formation during the transport of cholesterol to tissues. ApoB on the LDL particle acts as a ligand for LDL receptors in various cells throughout the body. High levels of ApoB can lead to plaques that cause vascular disease (atherosclerosis), leading to heart disease. There is considerable evidence that levels of ApoB are a better indicator of heart disease risk than total cholesterol or LDL (Contois et al, 2011; J. Clin. Lipid. 5 (4): 264-272).

There are two forms of ApoB (ApoB48 and ApoB100), with tissue regulated editing of ApoB48 and ApoB100 (reviewed in Davidson 2000; Ann. Rev. Nutr.; 20: 169-193). Editing is restricted to those transcripts expressed in the small intestine. This shorter version of the protein has a function specific to the small intestine. Editing results in a codon change creating an in frame stop codon leading to translation of a truncated protein, ApoB48. This stop codon results in the translation of a protein which lacks the carboxyl terminus which contains the protein's LDLR binding domain. The full protein ApoB100 which has nearly 4500 amino acid is present in VLDL and LDL. The main function of the full length liver expressed ApoB100 is as ligand for activation of the LDL-R. However editing results in a protein lacking this LDL-R binding region of the protein. This alters the function of the protein and the shorter ApoB48 protein as specific functions relative to the small intestine. ApoB48 is identical to the amino terminal 48% of ApoB100 (Knott et al., 1986; Nature 323 (6090): 734-8). The function of this isoform is in fat absorption of the small intestine and is involved in the synthesis, assembly and secretion of chylomicrons. These chylomicrons transport dietary lipids to tissues while the remaining chylomicrons along with associated residual lipids are in 2-3 hours taken up by the liver via the interaction of apolipoprotein E (ApoE) with lipoprotein receptors. It is the dominant ApoB protein in the small intestine of most mammals and the key protein in the exogenous pathway of lipoprotein metabolism.

Protein: ApoB Gene: APOB (Homo sapiens, chromosome 2, 21224301-21266945 [NCBI Reference Sequence: NC_(—)000002.11]; start site location: 21266817; strand: negative)

Gene Identification GeneID 338 HGNC 603 HPRD 00133 MIM 107730

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 2252 CGGTGGGGCGGCTCCTGGGCTGC 10 2329 CCTCGCGGCCCTGGCTGGCTGGGCG 46 2406 AACCGAGAAGGGCACTCAGCCCCG 88 2440 CGGCGCCCGCACCCCATTTATAGG 136 2451 GTCCAAAGGGCGCCTCCCGGGCC 195 2475 CGTCTTCAGTGCTCTGGCGCGGCC 341 2513 CACCGGAAGCTTCAGCCAGCGCTCGCTG 988 2552 CGAGTGGGAGGCGGCCAGGAGCAAGCCG 1281 2553 CGTACACTCACGGAAATGCTGTAAAG 2533 2576 CGTCACAGCCAATAATGAGCGTACGC 4862

Targeted Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 2252 CGGTGGGGCGGCTCCTGGGCTGC 10 2253 GGTGGGGCGGCTCCTGGGCT 11 2254 GTGGGGCGGCTCCTGGGCTG 12 2255 TGGGGCGGCTCCTGGGCTGC 13 2256 GGGGCGGCTCCTGGGCTGCG 14 2257 GGGCGGCTCCTGGGCTGCGG 15 2258 GGCGGCTCCTGGGCTGCGGC 16 2259 GCGGCTCCTGGGCTGCGGCC 17 2260 CGGCTCCTGGGCTGCGGCCT 18 2261 GGCTCCTGGGCTGCGGCCTG 19 2262 GCTCCTGGGCTGCGGCCTGG 20 2263 CTCCTGGGCTGCGGCCTGGC 21 2264 TCCTGGGCTGCGGCCTGGCC 22 2265 CCTGGGCTGCGGCCTGGCCT 23 2266 CTGGGCTGCGGCCTGGCCTC 24 2267 TGGGCTGCGGCCTGGCCTCG 25 2268 GGGCTGCGGCCTGGCCTCGG 26 2269 GGCTGCGGCCTGGCCTCGGC 27 2270 GCTGCGGCCTGGCCTCGGCC 28 2271 CTGCGGCCTGGCCTCGGCCT 29 2272 TGCGGCCTGGCCTCGGCCTC 30 2273 GCGGCCTGGCCTCGGCCTCG 31 2274 CGGCCTGGCCTCGGCCTCGC 32 2275 GGCCTGGCCTCGGCCTCGCG 33 2276 GCCTGGCCTCGGCCTCGCGG 34 2277 CCTGGCCTCGGCCTCGCGGC 35 2278 CTGGCCTCGGCCTCGCGGCC 36 2279 TGGCCTCGGCCTCGCGGCCC 37 2280 GGCCTCGGCCTCGCGGCCCT 38 2281 GCCTCGGCCTCGCGGCCCTG 39 2282 CCTCGGCCTCGCGGCCCTGG 40 2283 CTCGGCCTCGCGGCCCTGGC 41 2284 TCGGCCTCGCGGCCCTGGCT 42 2285 CGGCCTCGCGGCCCTGGCTG 43 2286 GGCCTCGCGGCCCTGGCTGG 44 2287 GCCTCGCGGCCCTGGCTGGC 45 2288 CCTCGCGGCCCTGGCTGGCT 46 2289 CTCGCGGCCCTGGCTGGCTG 47 2290 TCGCGGCCCTGGCTGGCTGG 48 2291 CGCGGCCCTGGCTGGCTGGG 49 2292 GCGGCCCTGGCTGGCTGGGC 50 2293 CGGCCCTGGCTGGCTGGGCG 51 2294 GGCCCTGGCTGGCTGGGCGG 52 2295 GCCCTGGCTGGCTGGGCGGG 53 2296 CCCTGGCTGGCTGGGCGGGC 54 2297 CCTGGCTGGCTGGGCGGGCT 55 2298 CTGGCTGGCTGGGCGGGCTC 56 2299 TGGCTGGCTGGGCGGGCTCC 57 2300 GGCTGGCTGGGCGGGCTCCT 58 2301 GCTGGCTGGGCGGGCTCCTC 59 2302 CTGGCTGGGCGGGCTCCTCA 60 2303 TGGCTGGGCGGGCTCCTCAG 61 2304 GGCTGGGCGGGCTCCTCAGC 62 2305 GCTGGGCGGGCTCCTCAGCG 63 2306 CTGGGCGGGCTCCTCAGCGG 64 2307 TGGGCGGGCTCCTCAGCGGC 65 2308 GGGCGGGCTCCTCAGCGGCA 66 2309 GGCGGGCTCCTCAGCGGCAG 67 2310 GCGGGCTCCTCAGCGGCAGC 68 2311 CGGGCTCCTCAGCGGCAGCA 69 2312 GGGCTCCTCAGCGGCAGCAA 70 2313 GGCTCCTCAGCGGCAGCAAC 71 2314 GCTCCTCAGCGGCAGCAACC 72 2315 CTCCTCAGCGGCAGCAACCG 73 2316 TCCTCAGCGGCAGCAACCGA 74 2317 CCTCAGCGGCAGCAACCGAG 75 2318 CTCAGCGGCAGCAACCGAGA 76 2319 TCAGCGGCAGCAACCGAGAA 77 2320 GCGGTGGGGCGGCTCCTGGG 9 2321 TGCGGTGGGGCGGCTCCTGG 8 2322 CTGCGGTGGGGCGGCTCCTG 7 2323 GCTGCGGTGGGGCGGCTCCT 6 2324 AGCTGCGGTGGGGCGGCTCC 5 2325 CAGCTGCGGTGGGGCGGCTC 4 2326 CCAGCTGCGGTGGGGCGGCT 3 2327 GCCAGCTGCGGTGGGGCGGC 2 2328 CGCCAGCTGCGGTGGGGCGG 1 2329 CCTCGCGGCCCTGGCTGGCTGGGCG 46 2330 CTCGCGGCCCTGGCTGGCTG 47 2331 TCGCGGCCCTGGCTGGCTGG 48 2332 CGCGGCCCTGGCTGGCTGGG 49 2333 GCGGCCCTGGCTGGCTGGGC 50 2334 CGGCCCTGGCTGGCTGGGCG 51 2335 GGCCCTGGCTGGCTGGGCGG 52 2336 GCCCTGGCTGGCTGGGCGGG 53 2337 CCCTGGCTGGCTGGGCGGGC 54 2338 CCTGGCTGGCTGGGCGGGCT 55 2339 CTGGCTGGCTGGGCGGGCTC 56 2340 TGGCTGGCTGGGCGGGCTCC 57 2341 GGCTGGCTGGGCGGGCTCCT 58 2342 GCTGGCTGGGCGGGCTCCTC 59 2343 CTGGCTGGGCGGGCTCCTCA 60 2344 TGGCTGGGCGGGCTCCTCAG 61 2345 GGCTGGGCGGGCTCCTCAGC 62 2346 GCTGGGCGGGCTCCTCAGCG 63 2347 CTGGGCGGGCTCCTCAGCGG 64 2348 TGGGCGGGCTCCTCAGCGGC 65 2349 GGGCGGGCTCCTCAGCGGCA 66 2350 GGCGGGCTCCTCAGCGGCAG 67 2351 GCGGGCTCCTCAGCGGCAGC 68 2352 CGGGCTCCTCAGCGGCAGCA 69 2353 GGGCTCCTCAGCGGCAGCAA 70 2354 GGCTCCTCAGCGGCAGCAAC 71 2355 GCTCCTCAGCGGCAGCAACC 72 2356 CTCCTCAGCGGCAGCAACCG 73 2357 TCCTCAGCGGCAGCAACCGA 74 2358 CCTCAGCGGCAGCAACCGAG 75 2359 CTCAGCGGCAGCAACCGAGA 76 2360 TCAGCGGCAGCAACCGAGAA 77 2361 GCCTCGCGGCCCTGGCTGGC 45 2362 GGCCTCGCGGCCCTGGCTGG 44 2363 CGGCCTCGCGGCCCTGGCTG 43 2364 TCGGCCTCGCGGCCCTGGCT 42 2365 CTCGGCCTCGCGGCCCTGGC 41 2366 CCTCGGCCTCGCGGCCCTGG 40 2367 GCCTCGGCCTCGCGGCCCTG 39 2368 GGCCTCGGCCTCGCGGCCCT 38 2369 TGGCCTCGGCCTCGCGGCCC 37 2370 CTGGCCTCGGCCTCGCGGCC 36 2371 CCTGGCCTCGGCCTCGCGGC 35 2372 GCCTGGCCTCGGCCTCGCGG 34 2373 GGCCTGGCCTCGGCCTCGCG 33 2374 CGGCCTGGCCTCGGCCTCGC 32 2375 GCGGCCTGGCCTCGGCCTCG 31 2376 TGCGGCCTGGCCTCGGCCTC 30 2377 CTGCGGCCTGGCCTCGGCCT 29 2378 GCTGCGGCCTGGCCTCGGCC 28 2379 GGCTGCGGCCTGGCCTCGGC 27 2380 GGGCTGCGGCCTGGCCTCGG 26 2381 TGGGCTGCGGCCTGGCCTCG 25 2382 CTGGGCTGCGGCCTGGCCTC 24 2383 CCTGGGCTGCGGCCTGGCCT 23 2384 TCCTGGGCTGCGGCCTGGCC 22 2385 CTCCTGGGCTGCGGCCTGGC 21 2386 GCTCCTGGGCTGCGGCCTGG 20 2387 GGCTCCTGGGCTGCGGCCTG 19 2388 CGGCTCCTGGGCTGCGGCCT 18 2389 GCGGCTCCTGGGCTGCGGCC 17 2390 GGCGGCTCCTGGGCTGCGGC 16 2391 GGGCGGCTCCTGGGCTGCGG 15 2392 GGGGCGGCTCCTGGGCTGCG 14 2393 TGGGGCGGCTCCTGGGCTGC 13 2394 GTGGGGCGGCTCCTGGGCTG 12 2395 GGTGGGGCGGCTCCTGGGCT 11 2396 CGGTGGGGCGGCTCCTGGGC 10 2397 GCGGTGGGGCGGCTCCTGGG 9 2398 TGCGGTGGGGCGGCTCCTGG 8 2399 CTGCGGTGGGGCGGCTCCTG 7 2400 GCTGCGGTGGGGCGGCTCCT 6 2401 AGCTGCGGTGGGGCGGCTCC 5 2402 CAGCTGCGGTGGGGCGGCTC 4 2403 CCAGCTGCGGTGGGGCGGCT 3 2404 GCCAGCTGCGGTGGGGCGGC 2 2405 CGCCAGCTGCGGTGGGGCGG 1 2406 AACCGAGAAGGGCACTCAGCCCCG 88 2407 ACCGAGAAGGGCACTCAGCC 89 2408 CCGAGAAGGGCACTCAGCCC 90 2409 CGAGAAGGGCACTCAGCCCC 91 2410 GAGAAGGGCACTCAGCCCCG 92 2411 AGAAGGGCACTCAGCCCCGC 93 2412 GAAGGGCACTCAGCCCCGCA 94 2413 AAGGGCACTCAGCCCCGCAG 95 2414 AGGGCACTCAGCCCCGCAGG 96 2415 GGGCACTCAGCCCCGCAGGT 97 2416 GGCACTCAGCCCCGCAGGTC 98 2417 GCACTCAGCCCCGCAGGTCC 99 2418 CACTCAGCCCCGCAGGTCCC 100 2419 ACTCAGCCCCGCAGGTCCCG 101 2420 CTCAGCCCCGCAGGTCCCGG 102 2421 TCAGCCCCGCAGGTCCCGGT 103 2422 CAGCCCCGCAGGTCCCGGTG 104 2423 AGCCCCGCAGGTCCCGGTGG 105 2424 GCCCCGCAGGTCCCGGTGGG 106 2425 CCCCGCAGGTCCCGGTGGGA 107 2426 CCCGCAGGTCCCGGTGGGAA 108 2427 CCGCAGGTCCCGGTGGGAAT 109 2428 CGCAGGTCCCGGTGGGAATG 110 2429 GCAGGTCCCGGTGGGAATGC 111 2430 CAGGTCCCGGTGGGAATGCG 112 2431 AGGTCCCGGTGGGAATGCGC 113 2432 GGTCCCGGTGGGAATGCGCG 114 2433 GTCCCGGTGGGAATGCGCGG 115 2434 TCCCGGTGGGAATGCGCGGC 116 2435 CCCGGTGGGAATGCGCGGCC 117 2436 CAACCGAGAAGGGCACTCAG 87 2437 GCAACCGAGAAGGGCACTCA 86 2438 AGCAACCGAGAAGGGCACTC 85 2439 CAGCAACCGAGAAGGGCACT 84 2440 CGGCGCCCGCACCCCATTTATAGG 136 2441 GGCGCCCGCACCCCATTTAT 137 2442 GCGCCCGCACCCCATTTATA 138 2443 CGCCCGCACCCCATTTATAG 139 2444 GCCCGCACCCCATTTATAGG 140 2445 CCCGCACCCCATTTATAGGA 141 2446 CCGCACCCCATTTATAGGAA 142 2447 CGCACCCCATTTATAGGAAG 143 2448 CCGGCGCCCGCACCCCATTT 135 2449 GCCGGCGCCCGCACCCCATT 134 2450 GGCCGGCGCCCGCACCCCAT 133 2451 GTCCAAAGGGCGCCTCCCGGGCC 195 2452 TCCAAAGGGCGCCTCCCGGG 196 2453 CCAAAGGGCGCCTCCCGGGC 197 2454 CAAAGGGCGCCTCCCGGGCC 198 2455 AAAGGGCGCCTCCCGGGCCT 199 2456 AAGGGCGCCTCCCGGGCCTG 200 2457 AGGGCGCCTCCCGGGCCTGA 201 2458 GGGCGCCTCCCGGGCCTGAC 202 2459 GGCGCCTCCCGGGCCTGACC 203 2460 GCGCCTCCCGGGCCTGACCT 204 2461 CGCCTCCCGGGCCTGACCTG 205 2462 GCCTCCCGGGCCTGACCTGT 206 2463 CCTCCCGGGCCTGACCTGTT 207 2464 CTCCCGGGCCTGACCTGTTT 208 2465 TCCCGGGCCTGACCTGTTTG 209 2466 CCCGGGCCTGACCTGTTTGC 210 2467 CCGGGCCTGACCTGTTTGCT 211 2468 CGGGCCTGACCTGTTTGCTT 212 2469 GGTCCAAAGGGCGCCTCCCG 194 2470 AGGTCCAAAGGGCGCCTCCC 193 2471 AAGGTCCAAAGGGCGCCTCC 192 2472 AAAGGTCCAAAGGGCGCCTC 191 2473 AAAAGGTCCAAAGGGCGCCT 190 2474 CAAAAGGTCCAAAGGGCGCC 189 2475 CGTCTTCAGTGCTCTGGCGCGGCC 341 2476 GTCTTCAGTGCTCTGGCGCG 342 2477 TCTTCAGTGCTCTGGCGCGG 343 2478 CTTCAGTGCTCTGGCGCGGC 344 2479 TTCAGTGCTCTGGCGCGGCC 345 2480 TCAGTGCTCTGGCGCGGCCC 346 2481 CAGTGCTCTGGCGCGGCCCT 347 2482 AGTGCTCTGGCGCGGCCCTT 348 2483 GTGCTCTGGCGCGGCCCTTC 349 2484 TGCTCTGGCGCGGCCCTTCC 350 2485 GCTCTGGCGCGGCCCTTCCT 351 2486 CTCTGGCGCGGCCCTTCCTG 352 2487 TCTGGCGCGGCCCTTCCTGT 353 2488 CTGGCGCGGCCCTTCCTGTG 354 2489 TGGCGCGGCCCTTCCTGTGT 355 2490 GGCGCGGCCCTTCCTGTGTC 356 2491 GCGCGGCCCTTCCTGTGTCT 357 2492 CGCGGCCCTTCCTGTGTCTC 358 2493 GCGGCCCTTCCTGTGTCTCA 359 2494 CGGCCCTTCCTGTGTCTCAG 360 2495 GCGTCTTCAGTGCTCTGGCG 340 2496 AGCGTCTTCAGTGCTCTGGC 339 2497 AAGCGTCTTCAGTGCTCTGG 338 2498 CAAGCGTCTTCAGTGCTCTG 337 2499 CCAAGCGTCTTCAGTGCTCT 336 2500 CCCAAGCGTCTTCAGTGCTC 335 2501 CCCCAAGCGTCTTCAGTGCT 334 2502 TCCCCAAGCGTCTTCAGTGC 333 2503 TTCCCCAAGCGTCTTCAGTG 332 2504 CTTCCCCAAGCGTCTTCAGT 331 2505 CCTTCCCCAAGCGTCTTCAG 330 2506 CCCTTCCCCAAGCGTCTTCA 329 2507 TCCCTTCCCCAAGCGTCTTC 328 2508 TTCCCTTCCCCAAGCGTCTT 327 2509 GTTCCCTTCCCCAAGCGTCT 326 2510 GGTTCCCTTCCCCAAGCGTC 325 2511 GGGTTCCCTTCCCCAAGCGT 324 2512 TGGGTTCCCTTCCCCAAGCG 323 2513 CACCGGAAGCTTCAGCCAGCGCTCGCTG 988 2514 ACCGGAAGCTTCAGCCAGCG 989 2515 CCGGAAGCTTCAGCCAGCGC 990 2516 CGGAAGCTTCAGCCAGCGCT 991 2517 GGAAGCTTCAGCCAGCGCTC 992 2518 GAAGCTTCAGCCAGCGCTCG 993 2519 AAGCTTCAGCCAGCGCTCGC 994 2520 AGCTTCAGCCAGCGCTCGCT 995 2521 GCTTCAGCCAGCGCTCGCTG 996 2522 CTTCAGCCAGCGCTCGCTGC 997 2523 TTCAGCCAGCGCTCGCTGCC 998 2524 TCAGCCAGCGCTCGCTGCCT 999 2525 CAGCCAGCGCTCGCTGCCTC 1000 2526 AGCCAGCGCTCGCTGCCTCT 1001 2527 GCCAGCGCTCGCTGCCTCTG 1002 2528 CCAGCGCTCGCTGCCTCTGC 1003 2529 CAGCGCTCGCTGCCTCTGCC 1004 2530 AGCGCTCGCTGCCTCTGCCC 1005 2531 GCGCTCGCTGCCTCTGCCCA 1006 2532 CGCTCGCTGCCTCTGCCCAG 1007 2533 GCTCGCTGCCTCTGCCCAGC 1008 2534 CTCGCTGCCTCTGCCCAGCT 1009 2535 TCGCTGCCTCTGCCCAGCTG 1010 2536 CGCTGCCTCTGCCCAGCTGG 1011 2537 CCACCGGAAGCTTCAGCCAG 987 2538 CCCACCGGAAGCTTCAGCCA 986 2539 TCCCACCGGAAGCTTCAGCC 985 2540 TTCCCACCGGAAGCTTCAGC 984 2541 TTTCCCACCGGAAGCTTCAG 983 2542 ATTTCCCACCGGAAGCTTCA 982 2543 CATTTCCCACCGGAAGCTTC 981 2544 CCATTTCCCACCGGAAGCTT 980 2545 CCCATTTCCCACCGGAAGCT 979 2546 GCCCATTTCCCACCGGAAGC 978 2547 TGCCCATTTCCCACCGGAAG 977 2548 CTGCCCATTTCCCACCGGAA 976 2549 ACTGCCCATTTCCCACCGGA 975 2550 CACTGCCCATTTCCCACCGG 974 2551 GCACTGCCCATTTCCCACCG 973 2552 CGAGTGGGAGGCGGCCAGGAGCAAGCCG 1281 2553 CGTACACTCACGGAAATGCTGTAAAG 2533 2554 GTACACTCACGGAAATGCTG 2534 2555 TACACTCACGGAAATGCTGT 2535 2556 ACACTCACGGAAATGCTGTA 2536 2557 CACTCACGGAAATGCTGTAA 2537 2558 GCGTACACTCACGGAAATGC 2532 2559 TGCGTACACTCACGGAAATG 2531 2560 TTGCGTACACTCACGGAAAT 2530 2561 CTTGCGTACACTCACGGAAA 2529 2562 ACTTGCGTACACTCACGGAA 2528 2563 GACTTGCGTACACTCACGGA 2527 2564 TGACTTGCGTACACTCACGG 2526 2565 CTGACTTGCGTACACTCACG 2525 2566 GCTGACTTGCGTACACTCAC 2524 2567 AGCTGACTTGCGTACACTCA 2523 2568 GAGCTGACTTGCGTACACTC 2522 2569 TGAGCTGACTTGCGTACACT 2521 2570 TTGAGCTGACTTGCGTACAC 2520 2571 GTTGAGCTGACTTGCGTACA 2519 2572 TGTTGAGCTGACTTGCGTAC 2518 2573 TTGTTGAGCTGACTTGCGTA 2517 2574 ATTGTTGAGCTGACTTGCGT 2516 2575 AATTGTTGAGCTGACTTGCG 2515 2576 CGTCACAGCCAATAATGAGCGTACGC 4862 2577 GTCACAGCCAATAATGAGCG 4863 2578 TCACAGCCAATAATGAGCGT 4864 2579 CACAGCCAATAATGAGCGTA 4865 2580 ACAGCCAATAATGAGCGTAC 4866 2581 CAGCCAATAATGAGCGTACG 4867 2582 AGCCAATAATGAGCGTACGC 4868 2583 GCCAATAATGAGCGTACGCA 4869 2584 CCAATAATGAGCGTACGCAA 4870 2585 ACGTCACAGCCAATAATGAG 4861 2586 GACGTCACAGCCAATAATGA 4860 2587 AGACGTCACAGCCAATAATG 4859 2588 CAGACGTCACAGCCAATAAT 4858 2589 TCAGACGTCACAGCCAATAA 4857 2590 ATCAGACGTCACAGCCAATA 4856 2591 AATCAGACGTCACAGCCAAT 4855 2592 TAATCAGACGTCACAGCCAA 4854 2593 ATAATCAGACGTCACAGCCA 4853 2594 CATAATCAGACGTCACAGCC 4852 2595 GCATAATCAGACGTCACAGC 4851 2596 GGCATAATCAGACGTCACAG 4850 2597 GGGCATAATCAGACGTCACA 4849 2598 AGGGCATAATCAGACGTCAC 4848 2599 AAGGGCATAATCAGACGTCA 4847 2600 GAAGGGCATAATCAGACGTC 4846

Hot Zones (Relative upstream location to gene start site)  1-600  700-1400 2450-2650 3450-3700 4600-5000

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11958) TGCATATGAAAGAAACCTATTCACATGGACCATATTACATTATAATCACA GTGTTTACTGCTTGACTACCATCTGCCTGGCCTAGCAAGGGTGTCAGTGA GGAAGAGAGGACAAGGGGTACCAATCTGTGAACTACACATGGTTCTTGCT CTCCCAGCTTCTCTCTCCCATTGGCAAGGCAACAGGTAAACACATGAAAA ATCAAATAATGCTATAAGAGAAAAATGTATTCAGGACAACAACAGGTTTG TATGAAGGCCTTTCATCATCGTTGTCCTACCTAGAAACTGAATGACAGGG AATCAGAGTCACAAGCTATGAAGTCTAACTGGGCTGTTCCCAGAGAAAGA TTCAGTGCAGTAGGTGGGGCTGCAGCCAGCCCTGGGTGGGTGGAAGGATG ACATCCACATAGGCAAGAGGGTGATAATTCACTTACGCAGCTCCTCACTG CACATTGAACCCTGCTGACTTCTGGCTTCTCTCCCGGGAGGAACTGCGAC TCAACATTCTGACCTTATCTCTTGGGTAGCAGAATGATGGAGAAGGAAAG TTTCTTTTTGCTTCTCGCAGGGGTTAATCATCCATCTGGAATGCCTACAT TTGGTTGACAATGGCTCACCCTATCATCTTCCTCCTGAACCATTCACCTA AATGTGCCATTTCTTTCCTGATAGTTCTCATTTGTGTGTGTGTGTGTGTG TGTGTGTGTGTGCACGTGCTCACACATGCATGCTGTCACTGGGTAAACAG GCCACCCTGGGCACAGTTCCATCTACAATGTTTGAAGTTTACTTTCCAGC TTCTGGGCATCATTTGCAATTATAATGCTGTCATAGGCAGAAACGAGATA GGCTAATTAATCGTTGTCAATACTGATCCCTATTTGCCAGATGAGATTTT GGAGCAGCATGGCTGGGAATAATTGGTATAGACTGTATTTCCTTGCTTTA TGTCACTGGAAATATTTATTTAAGCATCACGGTCGCTATGCATAAATATC CTGGAAAATGGGGTATAGCTGAATGGTGCAGATTCATTCATTCATATTCA GCAAATTATGTTCTAAGCACCTACTTCAGTATGTGAACAGCACTAAACTC AGAATATTGGTCTGCTGGGGTCCTTTATTAGCTTCCATGATTCCCTGAAC TTGGCCAAGACCCTTCTGGTCGGCTGCAGATAGGCACAATGGATAGTTTT GCTTCTAGATAATGTAACTGGGACATTCAGCATTATCTATCGCCTTGAAA TTCCTCTAGTCAGGTGGCTTTCTAATGGGTACCCAGAGCCCTATGACTAC CCAGATTGATGGTGCACCCAACAGGACTTTGCATTTATGAGCTGATAAGT CACAGTCACTAGCTGAGATTAATCTGTGTGACACCAGAATGTGTCTCTAT CTAAAGGAAAAGGGATGAAGGGTGATATCTTTGGTCACAAGTAATGTATT TCCATGTAGTCTTTGACAAAGGATCTAAGTGGATTTTGTAATTGAAGAAA AATCTATGCACTAATCTTTACAGCATTTCCGTGAGTGTACGCAAGTCAGC TCAACAATTCAACATTTGCTCTGTGGGGTTGTGCTAGACCCTGTCAGGGG ATAACTACTGCTGGCTGGGGCCCAGTTCAGGGAAGACTTGCCAAAGACCA TCAGGAAAAGAGGGAAGCTGAGTCTTAGGTTTCTTCCTTTAGAGATGGTG ACAGTCCTCTCACCACCTCCAAGCATCTCACAATGTTTCCCTGCCTCCAA GTCATCAAATTCATTTTTGATTCCTACTTCATAAAAATTACATTCTCCCA GCACTTTGGGAGGCCAAGGCGGGCAGATCATGAGGTCAGGAGTTCAAGAC CAGCCTGATCAACATGGTGAAACACCGTCTCTACTAAAAATACAAAAATT AGCTGGGCATAGTGGCACTCACCTGTTATCTCAGTTACTTGGGAGGCTAA GGCAGGAGAATCGCTTAAACCCGGGAGGCAGAGGTTGCAGTGAGCCGAGA TTGTACCACTACACTCCAGCCTGGGTGACAGAGGGAGACTCCATCTCAAA TAAATAAATTAAAAAAAAAAATATATATATATATGTATATATTCTCTATG GATGCTGACCATTGGACCCTGGTTTCATCTGCACGTAACAGAGTAAGCTT GGACTTGTGCTTGTAAATTAAAGCTCGACACCTCCTTTTGGCTTCTCTAT ACCTGAATATTCTTACTCACTCTCCTTAATGTGAATATGCATGGAAGCAG GACCATTTCCTCAAACACTAGCAGCAGCGAACCCTGTGGAAAGTCAGTCC ACATAGAATAATTCAAATAAAGTGTTCAGAGAAATGGGGTTTCAGAGCAA TTACTTTTTCCAGACCTTTCACAAATCAGTGGTGTAGGTATGACCAGCCT TGAGTTGAGACCTCTGTAATATCCATCTTTAATAACATTAATATGCTGTG GATGAGCAACTGATCACTGGAGGGAGTTTAGCTGCCCATAGGAGTTCATG GCTAATGACAATATCTGAATAAGGACAGGTGTGGAGCCCAGGTGCAGGAA GCAGGCGAAGGTCTTTCTGTGAGTCTCCTCTGAGGGAACTGGGTCTTTAT ACATAGTTACTGTTTCAGAATTGATCCTTCTGGAATCATCAGTCTTCACC AGTAGCTTGTTACATCTGGGGTTATCTCATAATTCAAACAAAGCTGACAA GTTGTAACAATGAGCACACACTGACTTCTGCAACAGGCGCTGTCCACTTC CCATCCGCACTCTACCGGCTTGCTCCTGGCCGCCTCCCACTCGCCTTCCT GGGTGGTCCCCCAGCAGTTATACCTACCTGGTTGTCGCCCCCTCTATCCT ACCACAATTGCTCACTAGCGGTTTCCTGCGTACACAGCTTGTCTCCCTAA CCAGAGTGGAGGTGCCTTGGGGACACAGCCAGGCTCAGACATTCACTCAG CTCATCATAGTGCCATCCCATCAATAACCCCTTCTGAGTGATCCTGGGTT AGTAAACCGAGTGTCCCTGAAATTCCACTACCGCTGATTCCCTCCAGCTG GGCAGAGGCAGCGAGCGCTGGCTGAAGCTTCCGGTGGGAAATGGGCAGTG CCTAGAAGAGAAGGAAACGATGCATGAGAAGGTTCCAGATGTCTATGAGG AACATGACGTGTCCTGTCCACTACTCTGCTTTTCCTCGTCCGCCTCCCCA CCACTGGAGGAAACCTAGAAGCTGGTGCAGGAAATCCTCCTCTCAACAAC CCAAGAACACTTTGCACAAGAGGGGTGCGCCCTCGGAGGTTGCTCTTCCC CAGAGGCCTCTCCTCGCTGGGGTTTCTTGAAGACAGATACTTGGACTCCT GCTGGGACCAGGCAGGCCACCCATCCTCAGGGGCAGTGACTGGTCACTCA CCAGACCTCCCTGCATCCCCCTTCTCTCTCCTCCCCCAGCACGGGCTGAA CCCCGCAGCCACAGATTCTGATCAGGATTAGGGTGTGGGTGCAAATCCAA GGTCCACCAAAATGGAAAAGAAGTAACCGATGGGAACACGTCTCCACCAA GACAGCGCTCAGGACTGGTTCTCCTCGTGGCTCCCAATTCAGTCCAGGAG AAGCAGAGATTTTGTCCCCATGGTGGGTCATCTGAAGAAGGCACCCCTGG TCAGGGCAGGCTTCTCAGACCCTGAGGCGCTGGCCATGGCCCCACTGAGA CACAGGAAGGGCCGCGCCAGAGCACTGAAGACGCTTGGGGAAGGGAACCC ACCTGGGACCCAGCCCCTGGTGGCTGCGGCTGCATCCCAGGTGGGCCCCC TCCCCGAGGCTCTTCAAGGCTCAAAGAGAAGCCAGTGTAGAAAAGCAAAC AGGTCAGGCCCGGGAGGCGCCCTTTGGACCTTTTGCAATCCTGGCGCTCT TGCAGCCTGGGCTTCCTATAAATGGGGTGCGGGCGCCGGCCGCGCATTCC CACCGGGACCTGCGGGGCTGAGTGCCCTTCTCGGTTGCTGCCGCTGAGGA GCCCGCCCAGCCAGCCAGGGCCGCGAGGCCGAGGCCAGGCCGCAGCCCAG GAGCCGCCCCACCGCAGCTGGCG ATG

10) IL17. Interleukin 17 is a cytokine is a potent mediator in delayed-type reactions by increasing chemokine production in various tissues to recruit monocytes and neutrophils to the site of inflammation. IL-17 is produced by T-helper cells and is induced by IL-23 which results in destructive tissue damage in delayed-type reactions. Interleukin 17 as a family functions as a proinflammatory cytokine that responds to the invasion of the immune system by extracellular pathogens and induces destruction of the pathogen's cellular matrix. Interleukin 17 acts synergistically with tumor necrosis factor and interleukin-1 (Chiricozzi et al., J Invest Dermatol. 2011 March; 131(3):677-87, Miossec et al., N. Engl. J. Med. 361 (9): 888-98). Most notably IL is involved in inducing many immune signaling molecules and mediating proinflammatory responses (e.g. allergic responses). IL-17 induces the production of many other cytokines (such as IL-6, G-CSF, GM-CSF, IL-1β, TGF-β, TNF-α), chemokines (including IL-8, GRO-α, and MCP-1), and prostaglandins (e.g., PGE2) from many cell types (fibroblasts, endothelial cells, epithelial cells, keratinocytes, and macrophages). The release of cytokines causes many functions, such as airway remodeling, a characteristic of IL-17 responses. The increased expression of chemokines attracts other cells including neutrophils. IL-17 function is also essential to a subset of CD4+ T-Cells called T helper 17 (Th17) cells. As a result of these roles, the IL-17 family has been linked to many immune/autoimmune related diseases including rheumatoid arthritis, psoriasis, ankylosing spondylitis asthma, lupus, allograft rejection and anti-tumor immunity (reviewed in Miossec and Kolls, Nature Reviews Drug Discovery 11, 763-776).

Protein: IL17 Gene: IL17A (Homo sapiens, chromosome 6, 52051185-52055436 [NCBI Reference Sequence: NC_(—)000006.11]; start site location: 52051230; strand: positive)

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 2601 CTTGTTTGTATCCGCATGGCTGTGCTC 4451 2616 CGAGACCGTTGAGGTGGAGTG 3148 2635 GGTCACTTACGTGGCGTGTCGC 107 2664 GACAAAATGTAGCGCTATCG 55

Target Shift Sequences Relative upstream location Sequence ID to gene No: Sequence (5′-3′) start site 2601 CTTGTTTGTATCCGCATGGCTGTGCTC 4451 2602 TTGTTTGTATCCGCATGGCT 4452 2603 TGTTTGTATCCGCATGGCTG 4453 2604 GTTTGTATCCGCATGGCTGT 4454 2605 TTTGTATCCGCATGGCTGTG 4455 2606 TTGTATCCGCATGGCTGTGC 4456

Gene Identification GeneID 3605 HGNC 5981 HPRD 04396 MIM 603149

2607 TGTATCCGCATGGCTGTGCT 4457 2608 GTATCCGCATGGCTGTGCTC 4458 2609 TATCCGCATGGCTGTGCTCC 4459 2610 ATCCGCATGGCTGTGCTCCT 4460 2611 TCCGCATGGCTGTGCTCCTG 4461 2612 CCGCATGGCTGTGCTCCTGA 4462 2613 CGCATGGCTGTGCTCCTGAG 4463 2614 GCTTGTTTGTATCCGCATGG 4450 2615 TGCTTGTTTGTATCCGCATG 4449 2616 CGAGACCGTTGAGGTGGAGTG 3148 2617 CCGAGACCGTTGAGGTGGAG 3147 2618 TCCGAGACCGTTGAGGTGGA 3146 2619 ATCCGAGACCGTTGAGGTGG 3145 2620 AATCCGAGACCGTTGAGGTG 3144 2621 CAATCCGAGACCGTTGAGGT 3143 2622 TCAATCCGAGACCGTTGAGG 3142 2623 TTCAATCCGAGACCGTTGAG 3141 2624 TTTCAATCCGAGACCGTTGA 3140 2625 GTTTCAATCCGAGACCGTTG 3139 2626 GGTTTCAATCCGAGACCGTT 3138 2627 AGGTTTCAATCCGAGACCGT 3137 2628 CAGGTTTCAATCCGAGACCG 3136 2629 TCAGGTTTCAATCCGAGACC 3135 2630 CTCAGGTTTCAATCCGAGAC 3134 2631 ACTCAGGTTTCAATCCGAGA 3133 2632 GACTCAGGTTTCAATCCGAG 3132 2633 TGACTCAGGTTTCAATCCGA 3131 2634 CTGACTCAGGTTTCAATCCG 3130 2635 GGTCACTTACGTGGCGTGTCGC 107 2636 GTCACTTACGTGGCGTGTCG 108 2637 TCACTTACGTGGCGTGTCGC 109 2638 CACTTACGTGGCGTGTCGCA 110 2639 ACTTACGTGGCGTGTCGCAG 111 2640 CTTACGTGGCGTGTCGCAGT 112 2641 TTACGTGGCGTGTCGCAGTG 113 2642 TACGTGGCGTGTCGCAGTGG 114 2643 ACGTGGCGTGTCGCAGTGGG 115 2644 CGTGGCGTGTCGCAGTGGGT 116 2645 GTGGCGTGTCGCAGTGGGTT 117 2646 TGGCGTGTCGCAGTGGGTTC 118 2647 GGCGTGTCGCAGTGGGTTCA 119 2648 GCGTGTCGCAGTGGGTTCAG 120 2649 CGTGTCGCAGTGGGTTCAGG 121 2650 GTGTCGCAGTGGGTTCAGGG 122 2651 TGTCGCAGTGGGTTCAGGGG 123 2652 GTCGCAGTGGGTTCAGGGGT 124 2653 TCGCAGTGGGTTCAGGGGTG 125 2654 CGCAGTGGGTTCAGGGGTGA 126 2655 TGGTCACTTACGTGGCGTGT 106 2656 GTGGTCACTTACGTGGCGTG 105 2657 TGTGGTCACTTACGTGGCGT 104 2658 CTGTGGTCACTTACGTGGCG 103 2659 TCTGTGGTCACTTACGTGGC 102 2660 TTCTGTGGTCACTTACGTGG 101 2661 CTTCTGTGGTCACTTACGTG 100 2662 CCTTCTGTGGTCACTTACGT 99 2663 TCCTTCTGTGGTCACTTACG 98 2664 GACAAAATGTAGCGCTATCG 55 2665 GGACAAAATGTAGCGCTATC 54

Hot Zones (Relative upstream location to gene start site)  1-150 2900-3250 4250-4600

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11959) CACTGTTCAGGACGGCCCTCAGGAGCACAGCCATGCGGATACAAACAAGC ATTATGCTGGAGAGGAACAATGATGCTATCAGGTTGGTACCAAGACTGAT ACTTCCTATTTAAGAAATCATGTTAATTATATAGGTATTAAGTTCTGGTT TTTGCAAGACAACATCGGTAACCTCAGAGATGAACTTCTAGTATGTAATA GTATTGATATCTTCAAATTCAGGGTAGATGATCTAGAGCTTGATAAACTG GGTCTAAATCTGAGTCTCAATTAAATCCTGATTTTATCAACTCCCCAAGG CCTTCCAGATGGCTCTCTATTGCAAATCTTCTCCTCTCTAAACATCACGT CTTCCTTTTAAATGTCTGTTTGACATTGCTCCTTGTGTGCTGGCCCATTT GCAAAGTATCAGCCCCTACATTTAAAATCCAAGAGAATTCTCTCCAACGA CTCATTTATTCTCTCTATATTAAGTGAAGATCAAGAGAGTAGTCTTGCCT GGTGATGTGCACAGCATTAATCAAAATGGCTCCAATCTTTTTGAACAGCC ACCATCTTTTATTAACTTTAAGCAGAAACAATGGCATGATGCCTCCTCCT ATACTGAGACCTTTGTTACTTTTATAAAATGGCTATAGAATATAAATTTA AAATATATTAATCTCTCAATGGTAACCTTACTACCTAAGCAAAAATTAAA ATTTATTTATTTTATCTTTTAGCAAGCCTTAATTTTCCACCACACTGAGA CAAAAGACTCAAAAATACTCTCTCCAGGACACAAAAGGAAAAATTTTTCT TTCCATCCCTTACATTATTCTCTATTGCAAGTAGAAAGAAATACTGTTTA ATGGCCTAATAATTTCAAGGAGCCTTGAAAATCATTTCTCTAAATCAAGA AGGGCAGAAACATTTTACCACTATCATCCCTTCAAATGCATGTTTGTCTC CTGGAAATCTTTCCCTGTCTCCAGTTGCATACTTGCCCCCACCCATTGAA GCTCTGAACTCACAACATGCAGTTCCCCATTGCTTATGGTGATCTGTTTC AAGCCCTCAGAGAAGCAAGATTGATAAACCTGGAAGACAATCACTAACAA AATAATCATCCCTAATTTACTACTCCCATTTGTTCTTTACTATATTCATC TCCAGAAAATCATAGATGATCAAAAGATTACCTGGTGCAAAACTCATTTT ATAAGACAGGAAACTAAGCCTCAAAGAGGTGAGGTACCCAAGGGCATGGG TAGTTAGTGGCAGATTCACAGTGAAAACAAAGGTGTCCTCTCATTCAGTG TTCTAGGCACTCTAATGCCCAGTCCAGCATGCACTCCACCTCAACGGTCT CGGATTGAAACCTGAGTCAGCTGGCTCTGCAGACAAATGCAGAGGAAGAG CCCTGCCCACTGTGAACACACTCTTACACTTCCAGCTGCTCCTCCAGGAG CCACAGGCTTCCAGCTCAAGCAAACGTCTGGGAAATCAGCCATACTCAAG GCTGCACACACCCAGCTCCAACCACCATGTCAAAATGATCCTTTTAACAC TTCTGTGAAAACCCATTTTGTTTCCCCATTTTATTTTAAAGCATTTCAAA GCATAGCACACAGGCTGAATTACATAGGCACAGGAGGCAAGACTAGGGAA AACAGCTAGAGTCAGCTTCTCCCCCTGCAACTTCAATCAAAATAGTTCAC AGTAAGCACATTCTCCCCTCCTTCTTCCTTGCCAGAAAGCAACAACAAGG AATCCTCCACTCCAGGGTGATCCTGAATAGGCTATAGCCTCATCACACTA AGTTCAAAAGGATCAGAATGCAGAGCCAAGGCACTAGACAACTCAGTAGG GTTATTGAGATGCAGGTAGGTTCTAAGAAGAATATAGAAAATTCTGAACT TCTCAAATAACATTACTCATACTGTCAATTAATCAAAATGTTGAACAGCT ATTACTTACTAGACACTATGCTAAGTGCTACAAAGACCAATAAGACTACT TGTCTTCAGGTAGCTTGTAGTCTAGCAAGAAATGAGCTACAAAGAAAACA AGAAAGTCAGTATGTAAATGAATGGCATGCAATACACTAGAAATATGCAT GAACAGCCCTGTATCAGAGAAGAACAAGCAACACAAAAGCAGTCTGTGAA GACCCATGTTGTATAAAGGGCCACATTAGGCACCATAGGAGTCACAAAGT TGAAGAACATACTAGCCTTGAAGGCTGACAGTAAAATCTAAACATGGAAA TAAACACATATCTTACCCATAGCTTTTAAAGCCAAATGAAACACATTACA AAATGAAAGAAAAGTTACTAGAGAGGTAGGTGTAAGCTACATGTGATGGA TGGAACTATTTATTTCAACTGAGGTAATCAGGCCAGACCTGTGAGATACA TTTAAACAGGATTTTGAAGGACAAGTAGCATTTTGATAACCTGAGGATGG GGAAGGGCATTTCAGAAACAATACCTTAACAAAGGGTGGGTAAGCTTTGA AAGAAGGCCTGGAGAAAAATCAGGACCCCATCATCCCAGGTCCTGGAGCA TGGTGGGGGGTAAGGCTAGAAAGGAAGTTGTACGAACTCAAATATTAATT CAAATGCTAAGAGGCTTACCCTTCATTCTGTATGCAAGCTAATGGCAGAA GAAAAGGCACAATTAGAGCCATGCTTTGAGAATCATTATTGAAAGCATGT CGAAGATGGTCTGAAGGAAGCAATTGGGAAAAGCAAGCATAGCTCATCCA AGTGGGTGAGAGTGTGAGTTAGAGGAAGCTTGGAAATTGGTGATGTGAGA GATGCTGCAGCTTCTGGGATTGCTGCCTGGTCGTGTGTAGAGGAGGGGCA GTAGGGCTCATTCTGAATCTTGTCTTGAAAAGCACATAGATAGTGATGCC AAAACCAGGACTACGGAAATCACTTGAAGCTGTATCCTACCTCCTCCTCC ATCTGTATCTGCTTCACCTATCAAGGATATCTACTATTGCCACTAAAATT CAGGTGCTTATGGCCTCCCTCATTCATCAGCCAGGGTTTATCTGGCCAGG AAAGAGAAGCCCCTTCAGGCATTTGCAACAGAGGGAGTTTAAGTCAGAGA ACTAGTCACCCTGGTAGTTGAGATTGCCATCAGCAAGAAGCTGTTACCAT TTGAAGGCTGCAGGGACAAAGGGAGTGAGCAGTCCTCTGGGAGACTGAGG AAGGAAGCTCCTGGCTTCTCCCCACTTTCCACTTTCCACTTCCCACTTTT ACTCATCTGTCCTCCAATTCCCTTTTGGCTGAGCCTAGCTGAAACCCAGC TGACAGGGGAGTTTGAGCAAGCAGCCTCCAGGGTCAGCCCTCTGAGTTAC AGGTAGAGCAGGACAGGGAGGAATGGATCTCAGGACAAACAGGTTCAGGA TCCGGCAAACTAATTTTACACTCCAGCCATTGAGTTGGAACTACTGGCCA GCCTCCCCCCGAGTTAGCATGTAGAATATGGGATACCAGCTGAGTGCCTG AGAGTTATCATTCACCTCAGTGGGGGTAGGGGCGGAGAAGGGTGACATAT AGCCAGCCACATCTATATCCACTGGCCCTTCCTTGTCCTAGTCCTCTGTA TTCCTGAGAAGGAACTATTCTCAAGGACCTGAGTCCAAGTTCATCTTACT TAGAGTACAGAGAAAAGAACCGCTAACTCCTTCTCTCTTTCCCCCATCAT GTCTCCTCTCCTTTCTAGTTCTCATCACTCTCTACTCCCCCCTGCCCCCC TTTTCTCCATCTCCATCACCTTTGTCCAGTCTCTATCCCCATTTTCAATT CCTTCCTCAAAACACCAAGTTGCTTGGTAGCATGCAGGGTTGGAACATGC CTTTAACAGAAAATCTCGTGTCTCTTGAACCTAGTTATTTATTCCTTGAG CAGAGTAGATATTCAACAAAAGAATTGTTAAATTCAATTAAATAGGATAT ATCTTATTATTAAATATTTTTTTCATTTTTTGTTTACTTATATGATGGGA ACTTGAGTAGTTTCCGGAATTGTCTCCACAACACCTGGCCAAGGAATCTG TGAGGAAAAGAAAGATCAAATGGAAAATCAAGGTACATGACACCAGAAGA CCTACATGTTACTTCAAACTTTTTCTTCCTCATGAACCATTAAAATAGAG CATAACTCTTCTGGCAGCTGTACATATGTTCATAAATACATGATATTGAC CCATAGCATAGCAGCTCTGCTCAGCTTCTAACAAGTAAGAATGAAAAGAG GACATGGTCTTTAGGAACATGAATTTCTGCCCTTCCCATTTTCCTTCAGA AGGAGAGATTCTTCTATGACCTCATTGGGGGCGGAAATTTTAACCAAAAT GGTGTCACCCCTGAACCCACTGCGACACGCCACGTAAGTGACCACAGAAG GAGAAAAGCCCTATAAAAAGAGAGACGATAGCGCTACATTTTGTCCATCT CATAGCAGGCACAAACTCATCCATCCCCAGTTGATTGGAAGAAACAACG A TG

11) MMP2. Matrix metalloproteinase-2 (MMP-2) is also known as 72 kDa type IV collagenase and gelatinase A is an enzyme that in humans is encoded by the MMP2 gene (Devarajan et al, 1992; J. Biol. Chem. 267 (35): 25228-32). The matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMPs are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. This gene encodes an enzyme which degrades type IV collagen, the major structural component of basement membranes. The enzyme plays a role in endometrial menstrual breakdown, regulation of vascularization and the inflammatory response. Mutations in the MMP2 gene are associated with Torg-Winchester syndrome, multicentric osteolysis and arthritis syndrome (Martignetti et al., 2001, Nat. Genet. 28 (3): 261-5).

Protein: MMP2 Gene: MMP2 (Homo sapiens, chromosome 16, 55513081-55540586 [NCBI Reference Sequence: NC_(—)000016.9]; start site location: 55513392; strand: positive)

Gene Identification GeneID 4313 HGNC 7166 HPRD 00386 MIM 120360

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 2666 GCTCCCTGGCCCCGCGCGTCGC 9 2732 CCGCGGCGCAGGGCTGCGCTCCGAG 85 2865 GCCGCCTGCTACTCCTGGCCTC 453 2869 GCGCACTCGGGCCCGCCCCTCTCTGCCC 361 2891 CGCTCCGAGGGTCCGCTGGCTCGG 101 3024 GTCCACCCTCAGTGCACGACCTCGT 478 3066 CACCGCCTGAGGAAGTCTGGATGC 239 3101 TGCCTCTCTCGCGATCTGGGCG 512 3131 GAGGGACGCCGGCTTGGCTAGGAC 618

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 2666 GCTCCCTGGCCCCGCGCGTCGC 7 2667 CTCCCTGGCCCCGCGCGTCG 8 2668 TCCCTGGCCCCGCGCGTCGC 9 2669 CCCTGGCCCCGCGCGTCGCC 10 2670 CCTGGCCCCGCGCGTCGCCC 11 2671 CTGGCCCCGCGCGTCGCCCG 12 2672 TGGCCCCGCGCGTCGCCCGG 13 2673 GGCCCCGCGCGTCGCCCGGG 14 2674 GCCCCGCGCGTCGCCCGGGG 15 2675 CCCCGCGCGTCGCCCGGGGG 16 2676 CCCGCGCGTCGCCCGGGGGT 17 2677 CCGCGCGTCGCCCGGGGGTC 18 2678 CGCGCGTCGCCCGGGGGTCG 19 2679 GCGCGTCGCCCGGGGGTCGC 20 2680 CGCGTCGCCCGGGGGTCGCT 21 2681 GCGTCGCCCGGGGGTCGCTG 22 2682 CGTCGCCCGGGGGTCGCTGG 23 2683 GTCGCCCGGGGGTCGCTGGC 24 2684 TCGCCCGGGGGTCGCTGGCT 25 2685 CGCCCGGGGGTCGCTGGCTC 26 2686 GCCCGGGGGTCGCTGGCTCG 27 2687 CCCGGGGGTCGCTGGCTCGG 28 2688 CCGGGGGTCGCTGGCTCGGT 29 2689 CGGGGGTCGCTGGCTCGGTG 30 2690 GGGGGTCGCTGGCTCGGTGC 31 2691 GGGGTCGCTGGCTCGGTGCG 32 2692 GGGTCGCTGGCTCGGTGCGT 33 2693 GGTCGCTGGCTCGGTGCGTG 34 2694 GTCGCTGGCTCGGTGCGTGT 35 2695 TCGCTGGCTCGGTGCGTGTG 36 2696 CGCTGGCTCGGTGCGTGTGG 37 2697 GCTGGCTCGGTGCGTGTGGC 38 2698 CTGGCTCGGTGCGTGTGGCC 39 2699 TGGCTCGGTGCGTGTGGCCG 40 2700 GGCTCGGTGCGTGTGGCCGC 41 2701 GCTCGGTGCGTGTGGCCGCC 42 2702 CTCGGTGCGTGTGGCCGCCT 43 2703 TCGGTGCGTGTGGCCGCCTC 44 2704 CGGTGCGTGTGGCCGCCTCG 45 2705 GGTGCGTGTGGCCGCCTCGC 46 2706 GTGCGTGTGGCCGCCTCGCC 47 2707 TGCGTGTGGCCGCCTCGCCG 48 2708 GCGTGTGGCCGCCTCGCCGC 49 2709 CGTGTGGCCGCCTCGCCGCC 50 2710 GTGTGGCCGCCTCGCCGCCT 51 2711 TGTGGCCGCCTCGCCGCCTG 52 2712 GTGGCCGCCTCGCCGCCTGG 53 2713 TGGCCGCCTCGCCGCCTGGT 54 2714 GGCCGCCTCGCCGCCTGGTT 55 2715 GCCGCCTCGCCGCCTGGTTG 56 2716 CCGCCTCGCCGCCTGGTTGG 57 2717 CGCCTCGCCGCCTGGTTGGA 58 2718 GCCTCGCCGCCTGGTTGGAG 59 2719 CCTCGCCGCCTGGTTGGAGC 60 2720 CTCGCCGCCTGGTTGGAGCC 61 2721 TCGCCGCCTGGTTGGAGCCT 62 2722 CGCCGCCTGGTTGGAGCCTG 63 2723 GCCGCCTGGTTGGAGCCTGC 64 2724 CCGCCTGGTTGGAGCCTGCT 65 2725 CGCCTGGTTGGAGCCTGCTC 66 2726 CGCTCCCTGGCCCCGCGCGT 6 2727 GCGCTCCCTGGCCCCGCGCG 5 2728 AGCGCTCCCTGGCCCCGCGC 4 2729 TAGCGCTCCCTGGCCCCGCG 3 2730 GTAGCGCTCCCTGGCCCCGC 2 2731 CGTAGCGCTCCCTGGCCCCG 1 2732 CCGCGGCGCAGGGCTGCGCTCCGAG 85 2733 CGCGGCGCAGGGCTGCGCTC 86 2734 GCGGCGCAGGGCTGCGCTCC 87 2735 CGGCGCAGGGCTGCGCTCCG 88 2736 GGCGCAGGGCTGCGCTCCGA 89 2737 GCGCAGGGCTGCGCTCCGAG 90 2738 CGCAGGGCTGCGCTCCGAGG 91 2739 GCAGGGCTGCGCTCCGAGGG 92 2740 CAGGGCTGCGCTCCGAGGGT 93 2741 AGGGCTGCGCTCCGAGGGTC 94 2742 GGGCTGCGCTCCGAGGGTCC 95 2743 GGCTGCGCTCCGAGGGTCCG 96 2744 GCTGCGCTCCGAGGGTCCGC 97 2745 CTGCGCTCCGAGGGTCCGCT 98 2746 TGCGCTCCGAGGGTCCGCTG 99 2747 GCGCTCCGAGGGTCCGCTGG 100 2748 CGCTCCGAGGGTCCGCTGGC 101 2749 GCTCCGAGGGTCCGCTGGCT 102 2750 CTCCGAGGGTCCGCTGGCTC 103 2751 TCCGAGGGTCCGCTGGCTCG 104 2752 CCGAGGGTCCGCTGGCTCGG 105 2753 CGAGGGTCCGCTGGCTCGGT 106 2754 GAGGGTCCGCTGGCTCGGTG 107 2755 AGGGTCCGCTGGCTCGGTGG 108 2756 GGGTCCGCTGGCTCGGTGGC 109 2757 GGTCCGCTGGCTCGGTGGCC 110 2758 GTCCGCTGGCTCGGTGGCCT 111 2759 TCCGCTGGCTCGGTGGCCTG 112 2760 CCGCTGGCTCGGTGGCCTGG 113 2761 CGCTGGCTCGGTGGCCTGGG 114 2762 GCTGGCTCGGTGGCCTGGGG 115 2763 CTGGCTCGGTGGCCTGGGGT 116 2764 TGGCTCGGTGGCCTGGGGTT 117 2765 GGCTCGGTGGCCTGGGGTTT 118 2766 GCTCGGTGGCCTGGGGTTTG 119 2767 CTCGGTGGCCTGGGGTTTGC 120 2768 TCGGTGGCCTGGGGTTTGCC 121 2769 CGGTGGCCTGGGGTTTGCCC 122 2770 GGTGGCCTGGGGTTTGCCCG 123 2771 GTGGCCTGGGGTTTGCCCGG 124 2772 TGGCCTGGGGTTTGCCCGGC 125 2773 GGCCTGGGGTTTGCCCGGCT 126 2774 GCCTGGGGTTTGCCCGGCTC 127 2775 CCTGGGGTTTGCCCGGCTCA 128 2776 CTGGGGTTTGCCCGGCTCAG 129 2777 TGGGGTTTGCCCGGCTCAGC 130 2778 GGGGTTTGCCCGGCTCAGCG 131 2779 GGGTTTGCCCGGCTCAGCGG 132 2780 GGTTTGCCCGGCTCAGCGGC 133 2781 GTTTGCCCGGCTCAGCGGCT 134 2782 TTTGCCCGGCTCAGCGGCTC 135 2783 TTGCCCGGCTCAGCGGCTCA 136 2784 TGCCCGGCTCAGCGGCTCAT 137 2785 GCCCGGCTCAGCGGCTCATG 138 2786 CCCGGCTCAGCGGCTCATGG 139 2787 CCGGCTCAGCGGCTCATGGT 140 2788 CGGCTCAGCGGCTCATGGTC 141 2789 GGCTCAGCGGCTCATGGTCC 142 2790 GCTCAGCGGCTCATGGTCCG 143 2791 CTCAGCGGCTCATGGTCCGG 144 2792 TCAGCGGCTCATGGTCCGGC 145 2793 CAGCGGCTCATGGTCCGGCC 146 2794 AGCGGCTCATGGTCCGGCCC 147 2795 GCGGCTCATGGTCCGGCCCC 148 2796 CGGCTCATGGTCCGGCCCCC 149 2797 GGCTCATGGTCCGGCCCCCG 150 2798 GCTCATGGTCCGGCCCCCGC 151 2799 CTCATGGTCCGGCCCCCGCG 152 2800 TCATGGTCCGGCCCCCGCGC 153 2801 CATGGTCCGGCCCCCGCGCC 154 2802 ATGGTCCGGCCCCCGCGCCC 155 2803 TGGTCCGGCCCCCGCGCCCC 156 2804 GGTCCGGCCCCCGCGCCCCA 157 2805 GTCCGGCCCCCGCGCCCCAG 158 2806 TCCGGCCCCCGCGCCCCAGC 159 2807 CCGGCCCCCGCGCCCCAGCC 160 2808 CGGCCCCCGCGCCCCAGCCC 161 2809 GGCCCCCGCGCCCCAGCCCC 162 2810 GCCCCCGCGCCCCAGCCCCC 163 2811 CCCCCGCGCCCCAGCCCCCG 164 2812 CCCCGCGCCCCAGCCCCCGC 165 2813 CCCGCGCCCCAGCCCCCGCC 166 2814 CCGCGCCCCAGCCCCCGCCG 167 2815 CGCGCCCCAGCCCCCGCCGC 168 2816 GCGCCCCAGCCCCCGCCGCC 169 2817 CGCCCCAGCCCCCGCCGCCG 170 2818 GCCCCAGCCCCCGCCGCCGC 171 2819 CCCCAGCCCCCGCCGCCGCC 172 2820 CCCAGCCCCCGCCGCCGCCG 173 2821 CCAGCCCCCGCCGCCGCCGC 174 2822 CAGCCCCCGCCGCCGCCGCC 175 2823 AGCCCCCGCCGCCGCCGCCG 176 2824 GCCCCCGCCGCCGCCGCCGC 177 2825 CCCCCGCCGCCGCCGCCGCC 178 2826 CCCCGCCGCCGCCGCCGCCG 179 2827 CCCGCCGCCGCCGCCGCCGC 180 2828 CCCCCGCCGCCGCCGCCGCC 181 2829 CCCCGCCGCCGCCGCCGCCG 182 2830 GCCCCCGCCGCCGCCGCCGC 183 2831 CCGCCGCCGCCGCCGCCGCA 184 2832 CGCCGCCGCCGCCGCCGCAG 185 2833 GCCGCCGCCGCCGCCGCAGG 186 2834 CCGCCGCCGCCGCCGCAGGT 187 2835 CGCCGCCGCCGCCGCAGGTC 188 2836 GCCGCCGCCGCCGCAGGTCC 189 2837 CCGCCGCCGCCGCAGGTCCT 190 2838 CGCCGCCGCCGCAGGTCCTG 191 2839 GCCGCCGCCGCAGGTCCTGG 192 2840 CCGCCGCCGCAGGTCCTGGC 193 2841 CGCCGCCGCAGGTCCTGGCA 194 2842 GCCGCCGCAGGTCCTGGCAA 195 2843 CCGCCGCAGGTCCTGGCAAT 196 2844 CGCCGCAGGTCCTGGCAATC 197 2845 GCCGCAGGTCCTGGCAATCC 198 2846 CCGCAGGTCCTGGCAATCCC 199 2847 CGCAGGTCCTGGCAATCCCT 200 2848 TCCGCGGCGCAGGGCTGCGC 84 2849 CTCCGCGGCGCAGGGCTGCG 83 2850 GCTCCGCGGCGCAGGGCTGC 82 2851 TGCTCCGCGGCGCAGGGCTG 81 2852 CTGCTCCGCGGCGCAGGGCT 80 2853 CCTGCTCCGCGGCGCAGGGC 79 2854 GCCTGCTCCGCGGCGCAGGG 78 2855 AGCCTGCTCCGCGGCGCAGG 77 2856 GAGCCTGCTCCGCGGCGCAG 76 2857 GGAGCCTGCTCCGCGGCGCA 75 2858 TGGAGCCTGCTCCGCGGCGC 74 2859 TTGGAGCCTGCTCCGCGGCG 73 2860 GTTGGAGCCTGCTCCGCGGC 72 2861 GGTTGGAGCCTGCTCCGCGG 71 2862 TGGTTGGAGCCTGCTCCGCG 70 2863 CTGGTTGGAGCCTGCTCCGC 69 2864 CCTGGTTGGAGCCTGCTCCG 68 2865 GCCGCCTGCTACTCCTGGCCTC 453 2866 CCGCCTGCTACTCCTGGCCT 454 2867 CGCCTGCTACTCCTGGCCTC 455 2868 GGCCGCCTGCTACTCCTGGC 452 2869 GCGCACTCGGGCCCGCCCCTCTCTGCCC 361 2870 CGCACTCGGGCCCGCCCCTC 362 2871 GCACTCGGGCCCGCCCCTCT 363 2872 CACTCGGGCCCGCCCCTCTC 364 2873 ACTCGGGCCCGCCCCTCTCT 365 2874 CTCGGGCCCGCCCCTCTCTG 366 2875 TCGGGCCCGCCCCTCTCTGC 367 2876 CGGGCCCGCCCCTCTCTGCC 368 2877 GGGCCCGCCCCTCTCTGCCC 369 2878 GGCCCGCCCCTCTCTGCCCC 370 2879 GCCCGCCCCTCTCTGCCCCA 371 2880 CCCGCCCCTCTCTGCCCCAC 372 2881 CCGCCCCTCTCTGCCCCACC 373 2882 CGCCCCTCTCTGCCCCACCC 374 2883 GGCGCACTCGGGCCCGCCCC 360 2884 GGGCGCACTCGGGCCCGCCC 359 2885 GGGGCGCACTCGGGCCCGCC 358 2886 GGGGGCGCACTCGGGCCCGC 357 2887 GGGGGGCGCACTCGGGCCCG 356 2888 CGGGGGGCGCACTCGGGCCC 355 2889 GCGGGGGGCGCACTCGGGCC 354 2890 GGCGGGGGGCGCACTCGGGC 353 2891 CGCTCCGAGGGTCCGCTGGCTCGG 101 2892 GCTCCGAGGGTCCGCTGGCT 102 2893 CTCCGAGGGTCCGCTGGCTC 103 2894 TCCGAGGGTCCGCTGGCTCG 104 2895 CCGAGGGTCCGCTGGCTCGG 105 2896 CGAGGGTCCGCTGGCTCGGT 106 2897 GAGGGTCCGCTGGCTCGGTG 107 2898 AGGGTCCGCTGGCTCGGTGG 108 2899 GGGTCCGCTGGCTCGGTGGC 109 2900 GGTCCGCTGGCTCGGTGGCC 110 2901 GTCCGCTGGCTCGGTGGCCT 111 2902 TCCGCTGGCTCGGTGGCCTG 112 2903 CCGCTGGCTCGGTGGCCTGG 113 2904 CGCTGGCTCGGTGGCCTGGG 114 2905 GCTGGCTCGGTGGCCTGGGG 115 2906 CTGGCTCGGTGGCCTGGGGT 116 2907 TGGCTCGGTGGCCTGGGGTT 117 2908 GGCTCGGTGGCCTGGGGTTT 118 2909 GCTCGGTGGCCTGGGGTTTG 119 2910 CTCGGTGGCCTGGGGTTTGC 120 2911 TCGGTGGCCTGGGGTTTGCC 121 2912 CGGTGGCCTGGGGTTTGCCC 122 2913 GGTGGCCTGGGGTTTGCCCG 123 2914 GTGGCCTGGGGTTTGCCCGG 124 2915 TGGCCTGGGGTTTGCCCGGC 125 2916 GGCCTGGGGTTTGCCCGGCT 126 2917 GCCTGGGGTTTGCCCGGCTC 127 2918 CCTGGGGTTTGCCCGGCTCA 128 2919 CTGGGGTTTGCCCGGCTCAG 129 2920 TGGGGTTTGCCCGGCTCAGC 130 2921 GGGGTTTGCCCGGCTCAGCG 131 2922 GGGTTTGCCCGGCTCAGCGG 132 2923 GGTTTGCCCGGCTCAGCGGC 133 2924 GTTTGCCCGGCTCAGCGGCT 134 2925 TTTGCCCGGCTCAGCGGCTC 135 2926 TTGCCCGGCTCAGCGGCTCA 136 2927 TGCCCGGCTCAGCGGCTCAT 137 2928 GCCCGGCTCAGCGGCTCATG 138 2929 CCCGGCTCAGCGGCTCATGG 139 2930 CCGGCTCAGCGGCTCATGGT 140 2931 CGGCTCAGCGGCTCATGGTC 141 2932 GGCTCAGCGGCTCATGGTCC 142 2933 GCTCAGCGGCTCATGGTCCG 143 2934 CTCAGCGGCTCATGGTCCGG 144 2935 TCAGCGGCTCATGGTCCGGC 145 2936 CAGCGGCTCATGGTCCGGCC 146 2937 AGCGGCTCATGGTCCGGCCC 147 2938 GCGGCTCATGGTCCGGCCCC 148 2939 CGGCTCATGGTCCGGCCCCC 149 2940 GGCTCATGGTCCGGCCCCCG 150 2941 GCTCATGGTCCGGCCCCCGC 151 2942 CTCATGGTCCGGCCCCCGCG 152 2943 TCATGGTCCGGCCCCCGCGC 153 2944 CATGGTCCGGCCCCCGCGCC 154 2945 ATGGTCCGGCCCCCGCGCCC 155 2946 TGGTCCGGCCCCCGCGCCCC 156 2947 GGTCCGGCCCCCGCGCCCCA 157 2948 GTCCGGCCCCCGCGCCCCAG 158 2949 TCCGGCCCCCGCGCCCCAGC 159 2950 CCGGCCCCCGCGCCCCAGCC 160 2951 CGGCCCCCGCGCCCCAGCCC 161 2952 GGCCCCCGCGCCCCAGCCCC 162 2953 GCCCCCGCGCCCCAGCCCCC 163 2954 CCCCCGCGCCCCAGCCCCCG 164 2955 CCCCGCGCCCCAGCCCCCGC 165 2956 CCCGCGCCCCAGCCCCCGCC 166 2957 CCGCGCCCCAGCCCCCGCCG 167 2958 CGCGCCCCAGCCCCCGCCGC 168 2959 GCGCCCCAGCCCCCGCCGCC 169 2960 CGCCCCAGCCCCCGCCGCCG 170 2961 GCCCCAGCCCCCGCCGCCGC 171 2962 CCCCAGCCCCCGCCGCCGCC 172 2963 CCCAGCCCCCGCCGCCGCCG 173 2964 CCAGCCCCCGCCGCCGCCGC 174 2965 CAGCCCCCGCCGCCGCCGCC 175 2966 AGCCCCCGCCGCCGCCGCCG 176 2967 GCCCCCGCCGCCGCCGCCGC 177 2968 CCCCCGCCGCCGCCGCCGCC 178 2969 CCCCGCCGCCGCCGCCGCCG 179 2970 CCCGCCGCCGCCGCCGCCGC 180 2971 CCGCCGCCGCCGCCGCCGCC 181 2972 CGCCGCCGCCGCCGCCGCCG 182 2973 GCCGCCGCCGCCGCCGCCGC 183 2974 CCGCCGCCGCCGCCGCCGCA 184 2975 CGCCGCCGCCGCCGCCGCAG 185 2976 GCCGCCGCCGCCGCCGCAGG 186 2977 CCGCCGCCGCCGCCGCAGGT 187 2978 CGCCGCCGCCGCCGCAGGTC 188 2979 GCCGCCGCCGCCGCAGGTCC 189 2980 CCGCCGCCGCCGCAGGTCCT 190 2981 CGCCGCCGCCGCAGGTCCTG 191 2982 GCCGCCGCCGCAGGTCCTGG 192 2983 CCGCCGCCGCAGGTCCTGGC 193 2984 CGCCGCCGCAGGTCCTGGCA 194 2985 GCCGCCGCAGGTCCTGGCAA 195 2986 CCGCCGCAGGTCCTGGCAAT 196 2987 CGCCGCAGGTCCTGGCAATC 197 2988 GCCGCAGGTCCTGGCAATCC 198 2989 CCGCAGGTCCTGGCAATCCC 199 2990 CGCAGGTCCTGGCAATCCCT 200 2991 GCGCTCCGAGGGTCCGCTGG 100 2992 TGCGCTCCGAGGGTCCGCTG 99 2993 CTGCGCTCCGAGGGTCCGCT 98 2994 GCTGCGCTCCGAGGGTCCGC 97 2995 GGCTGCGCTCCGAGGGTCCG 96 2996 GGGCTGCGCTCCGAGGGTCC 95 2997 AGGGCTGCGCTCCGAGGGTC 94 2998 CAGGGCTGCGCTCCGAGGGT 93 2999 GCAGGGCTGCGCTCCGAGGG 92 3000 CGCAGGGCTGCGCTCCGAGG 91 3001 GCGCAGGGCTGCGCTCCGAG 90 3002 GGCGCAGGGCTGCGCTCCGA 89 3003 CGGCGCAGGGCTGCGCTCCG 88 3004 GCGGCGCAGGGCTGCGCTCC 87 3005 CGCGGCGCAGGGCTGCGCTC 86 3006 CCGCGGCGCAGGGCTGCGCT 85 3007 TCCGCGGCGCAGGGCTGCGC 84 3008 CTCCGCGGCGCAGGGCTGCG 83 3009 GCTCCGCGGCGCAGGGCTGC 82 3010 TGCTCCGCGGCGCAGGGCTG 81 3011 CTGCTCCGCGGCGCAGGGCT 80 3012 CCTGCTCCGCGGCGCAGGGC 79 3013 GCCTGCTCCGCGGCGCAGGG 78 3014 AGCCTGCTCCGCGGCGCAGG 77 3015 GAGCCTGCTCCGCGGCGCAG 76 3016 GGAGCCTGCTCCGCGGCGCA 75 3017 TGGAGCCTGCTCCGCGGCGC 74 3018 TTGGAGCCTGCTCCGCGGCG 73 3019 GTTGGAGCCTGCTCCGCGGC 72 3020 GGTTGGAGCCTGCTCCGCGG 71 3021 TGGTTGGAGCCTGCTCCGCG 70 3022 CTGGTTGGAGCCTGCTCCGC 69 3023 CCTGGTTGGAGCCTGCTCCG 68 3024 GTCCACCCTCAGTGCACGACCTCGT 478 3025 TCCACCCTCAGTGCACGACC 479 3026 CCACCCTCAGTGCACGACCT 480 3027 CACCCTCAGTGCACGACCTC 481 3028 ACCCTCAGTGCACGACCTCG 482 3029 CCCTCAGTGCACGACCTCGT 483 3030 CCTCAGTGCACGACCTCGTC 484 3031 CTCAGTGCACGACCTCGTCA 485 3032 TCAGTGCACGACCTCGTCAC 486 3033 CAGTGCACGACCTCGTCACC 487 3034 AGTGCACGACCTCGTCACCC 488 3035 GTGCACGACCTCGTCACCCC 489 3036 TGCACGACCTCGTCACCCCA 490 3037 GCACGACCTCGTCACCCCAC 491 3038 CACGACCTCGTCACCCCACT 492 3039 ACGACCTCGTCACCCCACTT 493 3040 CGACCTCGTCACCCCACTTG 494 3041 GACCTCGTCACCCCACTTGC 495 3042 ACCTCGTCACCCCACTTGCC 496 3043 CCTCGTCACCCCACTTGCCT 497 3044 CTCGTCACCCCACTTGCCTC 498 3045 TCGTCACCCCACTTGCCTCT 499 3046 CGTCACCCCACTTGCCTCTC 500 3047 CGTCCACCCTCAGTGCACGA 477 3048 ACGTCCACCCTCAGTGCACG 476 3049 TACGTCCACCCTCAGTGCAC 475 3050 CTACGTCCACCCTCAGTGCA 474 3051 TCTACGTCCACCCTCAGTGC 473 3052 CTCTACGTCCACCCTCAGTG 472 3053 CCTCTACGTCCACCCTCAGT 471 3054 GCCTCTACGTCCACCCTCAG 470 3055 GGCCTCTACGTCCACCCTCA 469 3056 TGGCCTCTACGTCCACCCTC 468 3057 CTGGCCTCTACGTCCACCCT 467 3058 CCTGGCCTCTACGTCCACCC 466 3059 TCCTGGCCTCTACGTCCACC 465 3060 CTCCTGGCCTCTACGTCCAC 464 3061 ACTCCTGGCCTCTACGTCCA 463 3062 TACTCCTGGCCTCTACGTCC 462 3063 CTACTCCTGGCCTCTACGTC 461 3064 GCTACTCCTGGCCTCTACGT 460 3065 TGCTACTCCTGGCCTCTACG 459 3066 CACCGCCTGAGGAAGTCTGGATGC 256 3067 ACCGCCTGAGGAAGTCTGGA 257 3068 CCGCCTGAGGAAGTCTGGAT 258 3069 CGCCTGAGGAAGTCTGGATG 259 3070 CCACCGCCTGAGGAAGTCTG 255 3071 GCCACCGCCTGAGGAAGTCT 254 3072 AGCCACCGCCTGAGGAAGTC 253 3073 CAGCCACCGCCTGAGGAAGT 252 3074 CCAGCCACCGCCTGAGGAAG 251 3075 TCCAGCCACCGCCTGAGGAA 250 3076 CTCCAGCCACCGCCTGAGGA 249 3077 CCTCCAGCCACCGCCTGAGG 248 3078 GCCTCCAGCCACCGCCTGAG 247 3079 AGCCTCCAGCCACCGCCTGA 246 3080 CAGCCTCCAGCCACCGCCTG 245 3081 GCAGCCTCCAGCCACCGCCT 244 3082 CGCAGCCTCCAGCCACCGCC 243 3083 GCGCAGCCTCCAGCCACCGC 242 3084 TGCGCAGCCTCCAGCCACCG 241 3085 ATGCGCAGCCTCCAGCCACC 240 3086 GATGCGCAGCCTCCAGCCAC 239 3087 AGATGCGCAGCCTCCAGCCA 238 3088 CAGATGCGCAGCCTCCAGCC 237 3089 CCAGATGCGCAGCCTCCAGC 236 3090 CCCAGATGCGCAGCCTCCAG 235 3091 CCCCAGATGCGCAGCCTCCA 234 3092 GCCCCAGATGCGCAGCCTCC 233 3093 AGCCCCAGATGCGCAGCCTC 232 3094 AAGCCCCAGATGCGCAGCCT 231 3095 AAAGCCCCAGATGCGCAGCC 230 3096 TAAAGCCCCAGATGCGCAGC 229 3097 TTAAAGCCCCAGATGCGCAG 228 3098 TTTAAAGCCCCAGATGCGCA 227 3099 GTTTAAAGCCCCAGATGCGC 226 3100 TGTTTAAAGCCCCAGATGCG 225 3101 TGCCTCTCTCGCGATCTGGGCG 512 3102 GCCTCTCTCGCGATCTGGGC 513 3103 CCTCTCTCGCGATCTGGGCG 514 3104 CTCTCTCGCGATCTGGGCGC 515 3105 TCTCTCGCGATCTGGGCGCA 516 3106 CTCTCGCGATCTGGGCGCAC 517 3107 TCTCGCGATCTGGGCGCACA 518 3108 CTCGCGATCTGGGCGCACAG 519 3109 TCGCGATCTGGGCGCACAGC 520 3110 CGCGATCTGGGCGCACAGCC 521 3111 GCGATCTGGGCGCACAGCCT 522 3112 CGATCTGGGCGCACAGCCTC 523 3113 GATCTGGGCGCACAGCCTCA 524 3114 ATCTGGGCGCACAGCCTCAG 525 3115 TCTGGGCGCACAGCCTCAGA 526 3116 CTGGGCGCACAGCCTCAGAA 527 3117 TGGGCGCACAGCCTCAGAAC 528 3118 GGGCGCACAGCCTCAGAACC 529 3119 GGCGCACAGCCTCAGAACCC 530 3120 GCGCACAGCCTCAGAACCCC 531 3121 CGCACAGCCTCAGAACCCCC 532 3122 TTGCCTCTCTCGCGATCTGG 511 3123 CTTGCCTCTCTCGCGATCTG 510 3124 ACTTGCCTCTCTCGCGATCT 509 3125 CACTTGCCTCTCTCGCGATC 508 3126 CCACTTGCCTCTCTCGCGAT 507 3127 CCCACTTGCCTCTCTCGCGA 506 3128 CCCCACTTGCCTCTCTCGCG 505 3129 ACCCCACTTGCCTCTCTCGC 504 3130 CACCCCACTTGCCTCTCTCG 503 3131 GAGGGACGCCGGCTTGGCTAGGAC 618 3132 AGGGACGCCGGCTTGGCTAG 619 3133 GGGACGCCGGCTTGGCTAGG 620 3134 GGACGCCGGCTTGGCTAGGA 621 3135 GACGCCGGCTTGGCTAGGAC 622 3136 ACGCCGGCTTGGCTAGGACA 623 3137 CGCCGGCTTGGCTAGGACAC 624 3138 GCCGGCTTGGCTAGGACACC 625 3139 CCGGCTTGGCTAGGACACCC 626 3140 CGGCTTGGCTAGGACACCCT 627 3141 GGAGGGACGCCGGCTTGGCT 617 3142 AGGAGGGACGCCGGCTTGGC 616 3143 TAGGAGGGACGCCGGCTTGG 615 3144 CTAGGAGGGACGCCGGCTTG 614 3145 ACTAGGAGGGACGCCGGCTT 613 3146 TACTAGGAGGGACGCCGGCT 612 3147 CTACTAGGAGGGACGCCGGC 611 3148 ACTACTAGGAGGGACGCCGG 610 3149 TACTACTAGGAGGGACGCCG 609 3150 GTACTACTAGGAGGGACGCC 608 3151 GGTACTACTAGGAGGGACGC 607 3152 CGGTACTACTAGGAGGGACG 606 3153 GCGGTACTACTAGGAGGGAC 605

Hot Zones (Relative upstream location to gene start site)   1-1100 1250-3050 3950-4250

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11960) TAAGCTTGCTTGACGCAGGGTAGTCACAAACCTTCAATTTGCAAAAT TGCTATCTCTGCACAGCACAGTAGGGCAAAGTGTGAATAAAATGAGGTAA CCTGTACCTCCAGCTAAAGTCCCAGAATTAACTTTCCTTGGCTCCAGTGG ATTCACAAGCCGGTATCTGAATCATCACCTCACCAAGATGCCTGGATCTG CCTTTTAGCCCAGCTTGGGTCACATTGCCACTGTGGAGCCAGGAGGTGGG TCACATCTGCTGAATTCAGACCTAGAGTTGGGGAGAAGTAGCTTCCCAAT GGGAAACTAAGGGGCAGCTACTAAAAGTAGGAGGACAGGGTCTTGGGAAG GCTTACATGGCACACGGCCACTACCTCCCTCAATGCTCTGTCCCCTGCTT AGTGTCCCGCACTGTAATTTCTGCCTCTTCATCAATAAAACACCACCTTA ATGCCACTGTACCCCAGCACCAAAAACAGTGTTCATCAAAAACTTCCCGA ATAAATGACAGAATTCATGTCATATCGCGACGTCTTCTAATCACAGCCTG CGTAGTTTTCTGGGGCTGCTGTAAAAAAGCACTACAGACTGGGTGGCTTA CAACAGAAATTTATTCTCTCAGAGCTCTGAGACTAGAAGTCCAAAACCAA CATGTCAGCAGGGCCACGCTCCCTCTGAAGCCTCTGGGGGAAGAATTCCT TCTTGTCTCTTCTAGCTTCTGGGTTGCAGGCAACTCCTTGGGTTGCAGGC ATTGCTCCAATTTCTGCCTCCATGGTCACATGGAGTTCTTCTTGCTGTGT GTCTCTGTGTCCAAAGTTTTGTCTTCTTATCATGACAACAGGCTTTGGAT TAGAGCCCACTCATCTTAACTTGATTGTATCTGCAAAGACCCTATTTCCA TGTGAGGGCACAGTCACAGGCATTGGGACTTGAACATATCTTTTTGGCAA CACAATTATATCCACTAAACAGCATTTTTGCATCTATATTAAGAATGACT AAAAGATGCTGCAGTAACAAACATATCCCAAAAGCTCCATGGTTCGTTGC CTAAGAGTTTGTTTTTCACTTACGAAGTCTGTAGTGGGTCTGGTTGCTTT CATTTTACGACTTTGCTAGATCAACACAGGGCTCCCAGGGTTACCGTGGC AGGGGAAGAGAGATATACAGGAGTGCACAGGGGCCCTGGAGCATATTGCA TGTGCTCACCAACAAGCCCATTATCATCAGATACTGCCTGTTCTCTCCTC GTCCCCTGGGCCATTCTAGGCCCCAAGTGAGGGTTCTTGGATCTCACACA AGAAGGAATTTGAGGCAAGTCCATAAAGTGAAAGCAAGTTTATTAAGAAA GTAAGGGAATAAAAGAATGGCTATTTCATCGGCAGATCAGCCCCAAAAGC TGCTGGTTGCCCATTTTTATGGTTATTTCTTGATTATATGCTAAACAAGG GGTAAATTATTCATGCTTCTGCCTTTTAGGCCGTATAGGATAACTTCCTG ACGTTGCCATGGCATATGTAAACAGTCGTGGCGCTGGTGGGAGTGTGGCA GTAAGGCTGACCAGAGGTTTTTCTCATCATCATCTTGGTTTTGGTGGGTC TTGGCCAGCTTCTTTACTGCAACCTGTTTTATCAGCAAGGTCTTTATGAC CTGTATCTTGTGCCAGCCTCCTATCTCATTTCGTGATTAGGATTGCCTTA ATTTACTGGTAATGCAGGCCAGCAGGTCTTAGTCTAACCCCTATTCAAGA TGGAGTTGCTCTGGTTCCAATGCCTCTGACATAGTCATTGCTGGGCACAG AAGCCTCTCTCTCACTGAGTGCTGCTTCTGCAGCCATTGCCATCTCTGAA TGGGCACCAAGCCCCACACTGGAAACCGATACCCTTGGAATGCCAAAACC ACAGAGGCTGTGAAGAGGTACAGTCAAAGCTGCCCTCTGCCAGAGGAGCT CCAAAGCATGTTATGCAGACTCCAGAGACTTTCTGAAAAGGTTAAAACTC AAATTGGGCAAGTGTAAATGAACAACACCTAGTGAAGGGAGTCACATACA AGGCAATACAATAGAGAGTGGTGAGGACTGTGGCAAACCAAAGTATGTGC CTCATCTAAAGGGAGCAGTCACTACTCAACTTCAGCAAATTATTGCCATA TGGAAACTGGCATCCAGTATTGCCAGATTTTTGGGAAATTTTTTTAAAAA AAGAAAACCAGAAAGTCAGATTTTTACATGAAGTTTCCCAAATTTCAAAA TGCTGTTCAGGCTGGATTTAGCCCACAGGCCACGAGTTTGCAGCCCCTGC TTTAGTGAGATAACTTTTTCCATTTTCACTCTCAGCTCTCAGCTCTCCAA CTTGGCTCTCTGGCTATCCACAGGACGTGGACATGAGCCCAGTGGGGCTG GGCCAGGAGGCAATCCCCCTTCCCAACTGACCTCAGTCTCGCCCTCTCCA AAACAGCCAAGGTTTGTCACTGGGTCAGGCTGAAGGGCCTGGCTCCCTCC TGCGGGGCAAGGTCCCTCCCAAGAGGGTCCTTTAAAACTGACTCTGGAAA GTCAGAGCACACACCCACCAGACAAGCCTGAACTTGTCTGAAGCCCACTG AGACCCAAGCCGCAGAGACTTTTCTAGCTGTGATGATCAAGACATAATCG TGACCTCCAATGCCCCCCACAAGTATATTGCTCCTGATTCTTTCAGCCCC TGACCTTACTTCTCAAACTGTTCCCTGCTGACCCCCAGTCCTATCTGCCC CCTTCCTAGGCTGGTCCTTACTGACCCCTCCAGCTCCATCCCCTCACCCT GTGCCCCACCTTTTTCAGATAGAAAAAACTTTCTTCTCCAGTGCCTCTTG CTGTTTTTCATCTCTGGGCCATTGTCAATGTTCCCTAAAACATTCCCCAT ATTCCCCACCCAGCACTCCACCTCTTTAGCTCTTCAGGTCTCAGCTCAGA AGTCACTTCTTCCAGGAAGCCTTCCTTGATTGTCTTTACTAGTTTAGGGG CTGAAGTCAGGCGTTCCCAACAGCCTGCTGGAGTTCCCCATCACAGCTTA TCTCTCAACTGTCTTTCCTGAGAGAGGGAGAAGACATTCCTCAGAGACGG TTGTCACAGGGAGAACTTCAAAATTGGGATTCGACCTGAGAGGCCACATG GATTCTTGGCTTGGCGCAGGAAAGGATTCAAGAGTGAGTGGGGAATTCGT GGAACTGAGGGCTCCTCCCCTTTTTAGACCATATAGGGTAAACCTCCCCA CATTGCCATGGCATTTATAAACTGCCATGGCACTGGTGGGTGCTTCCTTT AACATGCTAATGCATTATAATTAGCGTAAAATGAGCAGTGAGGATGACCA GAGGTCGCTTTCTTTGCCATCTTGGTTTTGGCTGGCTTCTTCACTGCATA CTGTTTTATCAGTGGGGTCTTTGTGACCTCTATCTTATTAAACCAGTCTT GCCCAATTTCTATCTCATCCTGTGACCGAGAATGCGGACCCTCCTGGGAG TGCAGCCCAGCAGGTCTCAGCCTCATTTTACCCAGCCCCCTGTTCAAGAT GGAGTCGCTCTGGTTCCAACGTCTCTAACGCGGGGCCCCTGACTGCTCTA TTTCCCAAGGTGTATCTAGCATCTCGCACTATACGAGGCCAAGTTAAGGC TTACACATTTGCAGAAGGAAAGAGGTAAGGAAGCAACCTGGGACCTTCCA CTGTCTCTGTTTCCATCTCTCTCTTTCCATCTCTGTTCATCCCAGAATCT CTCTGTCCCTATCCCTAAATATCGAAAATTTCTGTCTCTGACCATCTATC ATTGTGGCTGATCATCTGTTTCTGACCATTCCTTCCCGTTCCTGACCCCA GGGAGTGCAGGGTGTCCTAGCCAAGCCGGCGTCCCTCCTAGTAGTACCGC TGCTCTCTAACCTCAGGACGTCAAGGGCCTAGAGCGACAGATGTTTCCCA GCAGGGGGTTCTGAGGCTGTGCGCCCAGATCGCGAGAGAGGCAAGTGGGG TGACGAGGTCGTGCACTGAGGGTGGACGTAGAGGCCAGGAGTAGCAGGCG GCCGGGGAAAAGAGGTGGAGAAAGGAAAAAAGAGGAGAAAAGTGGAGGAG GGCGAGTAGGGGGGTGGGGCAGAGAGGGGCGGGCCCGAGTGCGCCCCCCG CCCCCAGCCCCGCTCTGCCAGCTCCCTCCCAGCCCAGCCGGCTACATCTG GCGGCTGCCCTCCCTTGTTTCCGCTGCATCCAGACTTCCTCAGGCGGTGG CTGGAGGCTGCGCATCTGGGGCTTTAAACATACAAAGGGATTGCCAGGAC CTGCGGCGGCGGCGGCGGCGGCGGGGGCTGGGGCGCGGGGGCCGGACCAT GAGCCGCTGAGCCGGGCAAACCCCAGGCCACCGAGCCAGCGGACCCTCGG AGCGCAGCCCTGCGCCGCGGAGCAGGCTCCAACCAGGCGGCGAGGCGGCC ACACGCACCGAGCCAGCGACCCCCGGGCGACGCGCGGGGCCAGGGAGCGC TACG ATG

12) FAP. Fibroblast activation protein, alpha (FAP) also known as seprase or 170 kDa melanoma membrane-bound gelatinase is a protein that in humans is encoded by the FAP gene. FAP is a homodimeric integral membrane gelatinase belonging to the serine protease family with dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. FAP has been found to be overexpressed in stromal fibroblasts of solid tumors and epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas. This protein is thought to be involved in the control of fibroblast growth or epithelial-mesenchymal interactions during development, tissue repair, and epithelial carcinogenesis (reviewed by Chiri and Charugi, Am J Cancer Res 2011; 1(4):482-497). FAP expression is seen on activated stromal fibroblasts of more than 90% of all human carcinomas. Stromal fibroblasts play an important role in the development, growth and metastasis of carcinomas. It has been shown that targeting FAP inhibits stromagenesis and growth of tumor in mice. Sibrotuzumab a monoclonal antibody and small molecules against FAP are being developed (Edosada et al., J. Biol. Chem. 2006: 281, 7437-7444).

Protein: FAP Gene: FAP (Homo sapiens, chromosome 2, 163027200-163100045 [NCBI Reference Sequence: NC_(—)000002.11]; start site location: 163099837; strand: negative)

Gene Identification GeneID 2191 HGNC 3590 HPRD 02674 MIM 600403

Targeted Sequences Relative upstream Sequence location ID No: Sequence (5′-3′) to gene start site 3154 CAGAGCGTGGGTCACTGGATCT 39 3171 CACCAACATCTGCTTACGTTGAC 272 3177 TCCACGGACTTTTGAATACCGTGC 133

Target Shift Sequences Relative upstream Sequence location ID No: Sequence (5′-3′) to gene start site 3154 CAGAGCGTGGGTCACTGGATCT 39 3155 AGAGCGTGGGTCACTGGATC 40 3156 GAGCGTGGGTCACTGGATCT 41 3157 AGCGTGGGTCACTGGATCTG 42 3158 GCGTGGGTCACTGGATCTGT 43 3159 CGTGGGTCACTGGATCTGTG 44 3160 TCAGAGCGTGGGTCACTGGA 38 3161 TTCAGAGCGTGGGTCACTGG 37 3162 CTTCAGAGCGTGGGTCACTG 36 3163 TCTTCAGAGCGTGGGTCACT 35 3164 GTCTTCAGAGCGTGGGTCAC 34 3165 TGTCTTCAGAGCGTGGGTCA 33 3166 CTGTCTTCAGAGCGTGGGTC 32 3167 TCTGTCTTCAGAGCGTGGGT 31 3168 TTCTGTCTTCAGAGCGTGGG 30 3169 ATTCTGTCTTCAGAGCGTGG 29 3170 AATTCTGTCTTCAGAGCGTG 28 3171 CACCAACATCTGCTTACGTTGAC 272 3172 ACCAACATCTGCTTACGTTG 273 3173 CCAACATCTGCTTACGTTGA 274 3174 CAACATCTGCTTACGTTGAC 275 3175 ACACCAACATCTGCTTACGT 271 3176 TACACCAACATCTGCTTACG 270 3177 TCCACGGACTTTTGAATACCGTGC 133 3178 CCACGGACTTTTGAATACCG 134 3179 CACGGACTTTTGAATACCGT 135 3180 ACGGACTTTTGAATACCGTG 136 3181 CGGACTTTTGAATACCGTGC 137 3182 GGACTTTTGAATACCGTGCC 138 3183 GACTTTTGAATACCGTGCCA 139

Hot Zones (Relative upstream location to gene start site) 1-400

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11961) TACCACTCAAAAGTTATGGGACTTTGGGGAAGTTATTTAGATTTTGT GTGCATCCATGTCCTCATCTGTAAAATGAGGATAATAATAGTACGAATGT TCTGGGGATAAAAGGAGATAGCACGTGCAAGTGCTGAGAAAAAAACGTCA TGATCAATAAGAGTATTCAATAGACATGAACTAGTAGTAGTAATATTCTC TAATCTAAAAATGCTAGTGAAAAAACAATATGTGTTAACAAGTTATGTTA GTCTATAGGTGCTTACACATTCTATTTATACTATTTAAACAGTCTTCTTG TTGGTTAACCACTTCTAAAAAGATGTAGTATTTCCCTATTTAAATGACAA TGAAGCACTGATTTATCTTCTGAGTCTTTGTGTCCTCAGCACTCAAAGAA ATGGGTTGTGTCTCTGTTCTGTTTCTTTTCTAACCCCATTTTGATAGTTA ACCCTTTGGGTCTCCAAGGAGCACTTGCCCTAATTATGATAGGATAATGA AACACTGACATCTAATTATAACATTTATGAAGGTAGTAGATGCCAATTAC AAACTAGGAGGCATGTGGCTTTATATTCTTCCTTATGTAACAATTGTGGT TTTAGAAAAGAGATCAGATTCAAAAAATAGTTTGAGCTATATCATTTCCC ACTGATGCTATATTATGCCACTTCTTCCAGATCTATAACTATGTGATAGT TATTTTGAGCTTTGAAGACCTCAGGACTCTTTCTAGCTTGGAACTCAAAA ATCTCTTTGACGATGGGTATTTTGTTGAATCCTTTCATTTAAGCAAGTCC CTGGAGGAAGAGCTTTTAACCCAGGCACTATATAAATAATCATCGGATTA ATAGACCCCGATTAAAAAAAAACTGTAAACAAATTAATATTTTGAACACA GTCCTTGTAGAGTGAAATTGTGTTCTTTTGAGATATGTGTAACAAAGTAC TTTGAGGATGTGAACATCATTAATATTTTAGCCTTAATTATTTTACCCTC ACAACCTTAAGTGTCCCCCAACAGCAAATCAGTGAAATGTCAATTATAAT TTTAAAAAAAATTTATCACTTTGGAATAAAACTTTAGGAAGTATCACAAA GAAGCAATGTAAGGTGGTAACACTGGGTCCTGTTAAAATCTTGGGCAAGT TATCCAGTTTTTCTAGGCTGTTTCCTAATCATCAAAATGAGTTTTGGTAT GGATCAGATGAAATAATGCATTAAAATCACTTTGTAGATAACAGTAAACA ATAAATGTTTATTGAATCTGAGGAATCAAATGGGTAGGATGTTAGGAGCT GTTAGGCTCTCTAGAAGCAAATTTTTACTTTAAAGAGTATAAAATCAGGC TTATGTTTACTGCACTTGTATCCTTATTCCCCTTGTAACTTGTCCTTAAA TAATTTGTCATGGCTTTTGGTTAATAAACACATCTCTCTTTCATCTCCCC ACCATAAAATAAAAAGATAACATCCTTATGCTCTCAGCATGGTCTTACCT TCAGACTCTAGAAATACATAGCTGGATGTGTTTTCTGGGAAAACATATAA ATTAAAATCATTTTTGGCAGGTAAACATTGGCTACTAATAAATAGTTCTA GTAAGCTCTCCTCCTTATAACCTAAGGATTTATGTTATAGCTCACTTATC CAGTGGGCTTACCAGAATGCAGTCATATTCCAAAGTCAGCCTTACAAGGT CCCTCTCTGGAAAGAACTCAGTTGATGAGCCTCTACTCTCCTAATTGCTG CCCTTACATTTTTACATGACAAATCATCATCCTTTCTGGTTTTGCAGCAT TTTAATAGACCCAGAGTTGCTTTCTGAGAAAAGTCTCAGTACTAACAAAG GTTAGAACTAACAAAGGTTGAACTAAGGATGACATGCATGTGGTATGTGC CCTTTCCAGTCCTCTTCCCAGGTCCATGAAGAAATCACCAACCAGTGAGA ACACTCTTTCTACAGAACACAAACTCAGCTTCAGGCTATTTCTGAACAAG AGCTTTAGGAAGCAAAAGGAAACAAGTATGTAAGAACATTAAGAACATAC CTGCCATACTTATACTAGAATGTAAGTTCCTTAAGGGCACAGACTGTGTC TGATTCATTTTTGTTTTCTCAACAAAAATGTTGATGGGGTGTTAACTAAT TGTATAAAGTAGGAATAAGAGTCAGTTTCGTAAATGTTTTTTAAAAGTTG ACAAAAGTATTTTCATTTTGATTCCAAAATTAAAAAAAGCTAATAAAGAT ATTACAATATTTTAAAAATCCAAATTTTATGAGAGTTCTTGTCTGGATGA AAATTAGAATACATTCACATTATCTCAAACGAATGAACATGTGTGACTTT ATAAAAACAATACCTCCCTAAACCATGAATTCAGATGGAAAAACTCGACA TCTTTATTTCTGCAGTCAGTCTCATTTTTCTTAAAACAGTTCAAACTAGT AAGAATTTTCCAGAAGTTACAGCTTGACTCACCCAACCTTCCAAGGAAAA AACAAAAAAACTTAAACAGACATTGTTTCACTCTCATCATTTCCCACCCT TACTAATAGTGGCAACTTAAGTGTATCTTAAAGCACTCCAACCTCTTCAT AGAGCCTATTAAATGAGTATCTTGTGGACACCCACACACAGTCATAGAAT CCTAAGTGGTGCTCAGACCAGTCACATGTCAGTGCATTCTTAATTGCTAG AGCTAACATGCTCTCAGCATGGTCTTTTAATTACACCCTAATAATTTATT ATAGTTTCTCTCTACAATGTAAAGTCTTGGAAATCACCCACTAAAAAGTG CCTGTGTACTCTGGGGCTTTGGCAGGCTAGGGCAGAACTTCTGAGAACAC GGTGTGTTCCAGAGAAGACAATCAATCTGAGAGGACTTACACAGAAACAG TTCATTCAGGACCTGGCTGCTGGCTTTTATCTGAGATCTGAGGATTTCAC AATCACTTGGAGATACCTACAAGTGTATAGCACACCTTGGATATTACTCT TAATGATTACTTCATTTTGTAAAGAGGTGACTCCACCAACAGCAAAGGAG AGGGCCCAGCCCCAGCCACCAGGAATACAGTTCTCTGCCAGTAAGTGCCT AATGACTCATTTTCCTCAACAGAATTTTCATAAGGCTGGAATTCAGGGAG GGATGTCTGGAGAATGTCTGAAAGGAAGTTCACAAGCCACTGTCCTGCTC TTTGCTGGAGAAAGTGTCCCGTGGTAGCCAGAGAAGTTGACTAAGGCAAA CAGCAACATGTTTTGGTAACATTTCCCCATTACCTTTCATGTACAATCCA AGAAAGGTTGCCATGAAGTGTTTTAATCAGGTTGGGAACATTATAAACTT CGAAAAAAGAAAACCATTAGTGGAAAAATTAAGGACACAGTAGATTTAAC AACTGTGTTTACGTGGAACCACAAAATCTATCCAAGTGAATTGCATTAAA ACAGACAGAACACTCCAAGAAACTGTTGTATGTGTATTTTTTTTAATTCA GTCAACCATTTTACTAATCTGTCAAGATGACCAATTTCTTTGGAATTATG TAGATTTAGCCAAAATGAAATTATACATAAGATTTACTTTTCTTTTCAGA TGCTTTTTTATTTATTTTTAAATCTTTATAATTACTAGATGTTCTCCTCT CTCAGAAGATATTCTGAGAGGAAAGCAAAAATACCACTCTTGTAAAGCCA TTTCCATTCTTCCAAAGGTCTGCTGGTAAATTATTCTTACTGATCTTTCC ATCTTTCTAGCCTGTGCATACACACCTAACCCATACTAAATTTCACCAGA TGGCATTTTATTTCTTTAAAGTAAAGCAGCCGTGGGTTTAGACAGTTGAA TTTTTAAACTTCTGTATTTACTGAAAGTGCATATGGTGCTATATGGACAA AGAAATTGTGCTGAAAGAAAAACATTTCTGTCTGCAATACCTCATAATCT TCCAGAGGAAAAAAAAGTGCAGTTATATGGCACATTTCTCACAAAATCTT ATGTGGCTTCAATGTTCTTCCTCTGTTAAAAAGTAGATATATGTTTAATG TACAGACCTGCAAGTTTCATTATTTTAAATTCATCTTTTAGTGGCAAATA AAAATGTTATGCAAAACCCAATGACTTGCTAAAGTGATCCTTCAGTGAAT TCTAGAAGAAAATGCAACATAAACCTGAACTGGTAAAAAAGAAAAAATAA AAACCTCTGTATGTCAACGTAAGCAGATGTTGGTGTAGTTACAAGGATGA GAAGGCTATAAAACTTCCCTTGAGTCACTCACAGTTCATTTGAGGGCCAA GAACGCCCCCAAAATCTGTTTCTAATTTTACAGAAATCTTTTGAAACTTG GCACGGTATTCAAAAGTCCGTGGAAAGAAAAAAACCTTGTCCTGGCTTCA GCTTCCAACTACAAAGACAGACTTGGTCCTTTTCAACGGTTTTCACAGAT CCAGTGACCCACGCTCTGAAGACAGAATTAGCTAACTTTCAAAAACATCT GGAAAA ATG

13) P-Selectin. P-selectin is a protein that in humans is encoded by the SELP gene. P-selectin functions as a cell adhesion molecule (CAM) on the surfaces of activated endothelial cells that line the inner surface of blood vessels and activated platelets. In unactivated endothelial cells, it is stored in granules called Weibel-Palade bodies, and α-granules in unactivated platelets (McEver et al., 1989, J. Clin. Invest. 84 (1): 92-9). P-selectin is located on chromosome 1q21-q24, spans>50 kb and contains 17 exons in human. P-selectin is constitutively expressed on megakaryocytes (the precursor of platelets) and endothelial cells (Pan and McEver, 1998; J. Biol. Chem. 273 (16): 10058-67). The expression of P-selectin consists of two distinct mechanisms. One involves P-selectin synthesis by megakaryocytes and endothelial cells, and sorted into membranes of secretory granules until they are activated by agonists such as thrombin and translocated to the plasma membrane from granules. Second, an increased level of mRNA and protein is induced by inflammatory mediators such as tumor necrosis factor-a (TNF-a), LPS, interleukin-4 (IL-4) while TNF-alpha. Selectin-neutralizing monoclonal antibodies, recombinant soluble P-selectin glycoprotein ligand 1 and small-molecule inhibitors of selectins have been tested in clinical trials on patients with multiple trauma, cardiac indications and pediatricasthma, respectively (reviewed in Ley, 2003; Trends Mol. Med, 9 (6): 263-267).

Protein: P-selectin Gene: SELP (Homo sapiens, chromosome 1, 169558087-169599377 [NCBI Reference Sequence: NC_(—)000001.10]; start site location: 169599312; strand: negative)

Gene Identification GeneID 6403 HGNC 10721 HPRD 01433 MIM 173610

Targeted Sequences Relative upstream Sequence location to gene ID Sequence (5′-3′) start site 3184 TAGCTACGAATAAAGAAATTTGTAG 2694

Hot Zones (Relative upstream location to gene start site) 1550-1800 2650-2800 3100-3250

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11962) GTCAGGCTGGTCTTGACTCCTGACCTCAGGTGATCCACTCACCTTG GCCTCCCAAAGCGCTGGGATTATGGCATGAGCCACTGAGTCTGGCTGAAT GTTAGCTCTCTTGATGCTGTCCCATAAATCTTGTAGGCTTTCATCATTTC TTTTCATTCTTTTTTCTCCTCTCACTGTATATTTTCAAAAACCTGTCTTC AGTTCACAGATTCTTTCTTCTGCTTGATCAAGTCTGCTACTGGTGATTTC TACTGCATTTCTCACTTCATTCATTATATTTTTCAGCTCCAATTTCTTTT ATGATTTCAATCTTTCTGTTACATTTCTTATGTTGTGCATTTATTGTTTC TCTGATTTCACCAAATTGTTTCTCTGTGTTTGCTTCAAAGTAACTGAGCT TCTTTAAAAACAATTATCTTGAATCCATTGTCAGGCCATTTGTAGTACTC CATTTCTTTTGGGTCAGCTACTGGGAAATTATTGTGTTTCTTAGGTGGTG ATATTTTAATTTGGGTTTTCATGTTTCTTGCTGCCTTACACTGCTGTCTG AGCATCTGGTGGATCTGCCCCAATTTCAGGCTGTATGGGCTGACTTTGGT GGAGAAATACCTTCTTATGTGGAATAATGCGAGGATGCTGGCTGGGTGGG ATGCAAAAGTTCTGACTTCAGTAGGGGCAAAGCTGTGTGGTCTCCATGCA GATCTGTCAGCTGAGGTTGGTGTTAGTGAATACTACAGGGATCCTTAGAG GCCAACACTGTGGGTATCTACAGTGGCAATGAGGCTGTTGAGGTTTTCAA TTGTGACAAGTCCTCCATATCTCTTTTTTTCCCCACCTGGGAAGTCATGA CTGAGGACATCCCTCTTGGAATTAGGTCTAACTTGCAGGCCTGCTCCTGG TGGTGGTGACACTGGTGTCTGATGAACAGTGCCCATGGAGTGGCCAAGAG CCAAGGCCTGAAGCATGGGCATGCATGGAGGGACCACAGCACCAGATTCA ATTGTAGCAATGGTACCAGTGCCCAAGGCACAGGCATACTTACTATCACA TTGATAATGGTGTGTAAAATGCAGGTACTTATAAAGCAGCTAAGGAGCCA GGGACTTTACTGCATGCATACGCAGAGCTACAGTGGCTCCAGGATCCAGG GTGTGGGCTAGCTCTCCTTGGTGGCTGAGCTGGTGACTAGAGCATGGACA GGCACAGAGAAACCTTGACTCTAGGACCCAGGGTGTTCACTAGCTCACTA TAGTGGTGGCTCTGGTGTTGGAGGTGTGGGTGTGTGTAGTACAGCCTCAG AGACAGGGTCTGGAGCGCAGGTGTGCACATTACTACAGCAGCTCTGGAGT TGAGAATATGGGTTACCTTTCTACAGTGGCTGAACTAGTGTCTGGAGCAA AGACTTTCACAGAGAGAACTTGGCTTGGGGTCCCAGGGTGAGATCTAGTT CACAACAGCAGTGACTCCAGTGTCTGAGACATGAGGAGGTGCACTGCAGC CACAGAGCCACAGTCCAGAGTGTGAATATCTGTAGAGCAGCCACAACTTT TGGGGATCAGGAACACACATAGACTTGTGAGAGGTGGTAACCCTGGCCCC AGTCCTGGGGCAGTGCAACAATAGCTGCTTCTTGGTGAGGGGGTGTGAGG GGTAGTGCAACTGTGTTTCCCTTTTTAGCATCCTGCTATGGGAATGGCTG TTGGATAAAAGATGCCAGTGTCCTCTGTGGAGCAGGACACTGGGGGCCTC AGTGGCTCTGTGTCACATGACTGACACAGATAGCCTACAAATTTCTTTAT TCGTAGCTATCTCCTGGTGTCTCATATATGCCAGTCTCACCGGTGATTCT TCTACATGGATATTCTTTCTTTTCTCCATTGTGTTGTTCCAAATTCTTTA ACAGGCTCTTGAGCCCCATCCCCCAACTCCCCACCCTTGTGAGGGCTATT TTGGTTTGTGTATAACTGTCTATGTTTGTTTTTTTGTTGGGGCATAAGGC TGACATCTCCTACTCCACCATCTTGCTAATGTCACCTGCATAGGAATCTT TTTATGCTTTCCTTATATTCACTAAAATTTAACAATATCAAACTTAAAAA CATATGATCAATTGAACTTATTAATATCAAACTTATTATAAATAAGAAAC TACCAGGCTGGGCATGGTGGCTCATGCCTGTAATCCCAACATTTTGGGAG GCTGAGGTGAAAGGATCACTTGAGCCCAGGAATTCAAGACCAGCCTGGGA AATATAGAGAGACCCTATCTCTAGAGATTTTTTTTTTTAATTAGCCAGTA GTGATGGCACACATCTATAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAG AATTGCTTGAGCCCAGGAGGTCAAGGCTGGAGCAAGCAGTAATCATGCCA CTGCACTCCAGCCTGGGCCGCAGAGTGAGACCCTGTCTCAAAAAAAGAAC CTACTAGTCTACATACCACACTTCCTCATCCCCATCTGAGACTATATATA TTTTTTCTAACATGAGGCAATGCCAAAAAGAGGGGCTGGTGAGTGAAAGT AAGAACAGAAAGACATGGAGGCAAGTCTTATAGAATAATAGCCAACACTT AAACTTACACTTAACAGCGTGATAGGTATTGTTCCAAACACATTAAATTC ATTTAATGGTCCTTACATGTCTATGTATTTGGTGATTATTATCCTTATTA TTCACATTGCTGAGTGTATTATTCTGTTCTCATGATGCTGATAGAGACAT ACCCGAGACTGGATAACTTATTAAAAAAAAAAAGGTTTAATGGACTCACA GTTCCACGTGGATGGGGAGTCCTCACAATCATGGTAGAAAGCAAAAGACA CGTCTTACATGGCAGCAGGGAAGAGAGAGAAATGAGAACCAAACAAAAGG GGTTTCCCCTTATAAAACCATCAGCTCTCATGCGACTTATTCACTACCAT GAGAACAGTATGGGGGAAACCACCCCCATGATTCAATGATCTACCAGGTG CCTCCCACAACCTGTGGGAATTATGGGAGCTACAATTCCAGATGAGATTT GGGTGGGGACACAGCCAAACCACATCACTGAGGAAACTGAGTTATAGGGA GATTAGTAACGCCCAACACAGCTGGTAGGTGGTGGAGCCAGGCAGTCTGA CTCTAGGGTCTGGACTCTGAACTGCATCATGCTGCCAAGAAGTTCCTCAT TTTTTCCTCTCTCTAAGTTTCCCTTATTCCCCTACAGTCATTCCTTCAAC AGCATTTCCTTCACCATCTTTTCTACTTCTACTATATAATTAATTTTTTC TTCTTGGTCCCAAATTCCAACGTGCAAATGCAGCCTTATATACCCTAATT CATCTTTACCTTTAGACTTTCTTCCAATGTTTCTACTTCATTCCATTTTA AATTTATCCATGAGATGCCTATTTACAAGCTGTAACCATCATGAAGTGAA TGAAGAATAATACCTACTACTGTACAATAGAATTCCAAGAGTATAAATAG GAGTTATGGCTTTCTGACTTGAAACTAAATACTTGATACTTGATTTTGCT GTCTGAGATCAATCTGAAAAGTAATAATAATCACTAACATTTGTTGAGCA TCAATTGTGGGCCAAGTGTCATTTCAATCACTCTGTACATATTAACTCAT TTCATCCTACAACAACCCGGTGAGGCAAGTTCTGTTATTCTGTTTTACAG TTGAGGAAACAGAGGCATAGAGAGCTTAAGTAGTTTGCCCAGTAGATAGC CAGAAGAGGAGCCAGGATGGGTCTCGGGCAGTTTAACAGCACAGCTGAAG TCTTAACCACTATGCCAACAGCTTTTTGGTCCTACACATCCCATGGGAAG AGGAAAATAAAAAGGTATCTATTTGTATACCTTTTTATTTCTGATATAAG AAGCAGAATTCCTTTCACATGACCTATGTCTATTTAATACGTCATTTTGA AACTTACCAATAAAATTTCCCAAGCGCCAGAAAACTGTTAGTGGCTTTTT CCATTTCTCTCTATTTTTTTTTGTGCTACTAATTTTGCTTCTTTCCCTCA GAAGGCTGCCGGAATAGTAAACATTCACTGACATGTCATAATTACTGGAA AATGGGCACTGGAAAATCACATTGTAATTAATTCAAAGCATGTTTTCCAA ATGTACTACTTTAAATTGGAGCTTATATCATAATCCAAGGAAACCTTTGT GTGTGTACTGTTCCCACATTGCTCAGCCTGGGATATCCAGGAGTAATTCA CCTTGCGCCTGCCTCCAGACCATCTTCCATGGAAGGGGGTGACCCCTTGC CTCTTGGCAACCACTATTTCTAAGCTGCCAACATTACTCTTGCATTATCA ACATTCTAACTTCATGGGAAGGGCTGTGGTGAGTTTCTGGAATGTGAATA GGAAGTTGTTTTTCTAAACAGCCTGACACTGAGGGGAGGCAGTGAGACTG TAAGCAGTCTGGGTTGGGCAGAAGGCAGAAAACCAGCAGAGTCACAGAGG AG ATG

14) IL-6. Interleukin 6 (IL-6) acts as both a pro-inflammatory and anti-inflammatory cytokine IL-6 is secreted by T cells and macrophages to stimulate immune response, e.g. during infection and after trauma, especially burns or other tissue damage leading to inflammation. IL-6 also plays a role in fighting infection, as IL-6 has been shown in mice to be required for resistance against bacterium Streptococcus pneumoniae. IL-6 is relevant to many diseases such as diabetes, atherosclerosis, depression, Alzheimer's Disease, systemic lupus erythematosus, multiple myeloma, prostate cancer, behcet's disease,[22] and rheumatoid arthritis (Kishimoto, International Immunology, Vol. 22, No. 5, pp. 347-352). IL-6 is also considered a myokine, a cytokine produced from muscle, and is elevated in response to muscle contraction. It is significantly elevated with exercise, and precedes the appearance of other cytokines in the circulation. During exercise, it is thought to act in a hormone-like manner to mobilize extracellular substrates and/or augment substrate delivery. Additionally, osteoblasts secrete IL-6 to stimulate osteoclast formation. Smooth muscle cells in the tunica media of many blood vessels also produce IL-6 as a pro-inflammatory cytokine IL-6's role as an anti-inflammatory cytokine is mediated through its inhibitory effects on TNF-alpha and IL-1, and activation of IL-1ra and IL-10.

Advanced/metastatic cancer patients have higher levels of IL-6 in their blood. IL-6 is responsible for stimulating acute phase protein synthesis, as well as the production of neutrophils in the bone marrow. It supports the growth of B cells and is antagonistic to regulatory T cells. Therefore there is interest in developing anti-IL-6 agents as therapy against many of these diseases (reviewed in Barton, Expert Opin. Ther. Targets 9 (4): 737-752).

Protein: IL-6 Gene: IL-6 (Homo sapiens, chromosome 7, 22766766-22771621 [NCBI Reference Sequence: NC_(—)000007.13]; start site location: 22766882; strand: positive)

Gene Identification GeneID 3569 HGNC 6018 HPRD 00970 MIM 147620

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 3185 CACCGCGTGGCTTCTGCCACTTTC 723 3206 TACGGACGCAGGCACGGCTCTAG 1117 3226 CAGCTCCGCAGCCGTGCACTGTG 1722 3255 CTTCACCGATTGTCTAAACAGAGAC 1525 3256 IL6_1 TTCGTTCCCGGTGGGCTCGAGGGC 35 3276 TGCTTCCGCGTCGGCACCCAAG 1150

Target Shift Sequences Relative upstream location to Sequence ID gene start No: Sequence (5′-3′) site 3185 CACCGCGTGGCTTCTGCCACTTTC 723 3186 ACCGCGTGGCTTCTGCCACT 724 3187 CCGCGTGGCTTCTGCCACTT 725 3188 CGCGTGGCTTCTGCCACTTT 726 3189 GCGTGGCTTCTGCCACTTTC 727 3190 CGTGGCTTCTGCCACTTTCT 728 3191 CCACCGCGTGGCTTCTGCCA 722 3192 GCCACCGCGTGGCTTCTGCC 721 3193 TGCCACCGCGTGGCTTCTGC 720 3194 TTGCCACCGCGTGGCTTCTG 719 3195 TTTGCCACCGCGTGGCTTCT 718 3196 TTTTGCCACCGCGTGGCTTC 717 3197 TTTTTGCCACCGCGTGGCTT 716 3198 CTTTTTGCCACCGCGTGGCT 715 3199 CCTTTTTGCCACCGCGTGGC 714 3200 TCCTTTTTGCCACCGCGTGG 713 3201 CTCCTTTTTGCCACCGCGTG 712 3202 ACTCCTTTTTGCCACCGCGT 711 3203 GACTCCTTTTTGCCACCGCG 710 3204 TGACTCCTTTTTGCCACCGC 709 3205 GTGACTCCTTTTTGCCACCG 708 3206 TACGGACGCAGGCACGGCTCTAG 1117 3207 ACGGACGCAGGCACGGCTCT 1118 3208 CGGACGCAGGCACGGCTCTA 1119 3209 GGACGCAGGCACGGCTCTAG 1120 3210 GACGCAGGCACGGCTCTAGG 1121 3211 ACGCAGGCACGGCTCTAGGC 1122 3212 CGCAGGCACGGCTCTAGGCT 1123 3213 GCAGGCACGGCTCTAGGCTC 1124 3214 CAGGCACGGCTCTAGGCTCT 1125 3215 AGGCACGGCTCTAGGCTCTG 1126 3216 GGCACGGCTCTAGGCTCTGA 1127 3217 GCACGGCTCTAGGCTCTGAA 1128 3218 CACGGCTCTAGGCTCTGAAT 1129 3219 ACGGCTCTAGGCTCTGAATC 1130 3220 CGGCTCTAGGCTCTGAATCT 1131 3221 CTACGGACGCAGGCACGGCT 1116 3222 ACTACGGACGCAGGCACGGC 1115 3223 AACTACGGACGCAGGCACGG 1114 3224 AAACTACGGACGCAGGCACG 1113 3225 GAAACTACGGACGCAGGCAC 1112 3226 CAGCTCCGCAGCCGTGCACTGTG 1700 3227 AGCTCCGCAGCCGTGCACTG 1701 3228 GCTCCGCAGCCGTGCACTGT 1702 3229 CTCCGCAGCCGTGCACTGTG 1703 3230 TCCGCAGCCGTGCACTGTGA 1704 3231 CCGCAGCCGTGCACTGTGAT 1705 3232 CGCAGCCGTGCACTGTGATC 1706 3233 GCAGCCGTGCACTGTGATCC 1707 3234 CAGCCGTGCACTGTGATCCG 1708 3235 AGCCGTGCACTGTGATCCGT 1709 3236 GCCGTGCACTGTGATCCGTC 1710 3237 CCGTGCACTGTGATCCGTCT 1711 3238 CGTGCACTGTGATCCGTCTA 1712 3239 GTGCACTGTGATCCGTCTAT 1713 3240 TGCACTGTGATCCGTCTATG 1714 3241 GCACTGTGATCCGTCTATGT 1715 3242 CACTGTGATCCGTCTATGTA 1716 3243 CCAGCTCCGCAGCCGTGCAC 1699 3244 CCCAGCTCCGCAGCCGTGCA 1698 3245 TCCCAGCTCCGCAGCCGTGC 1697 3246 CTCCCAGCTCCGCAGCCGTG 1696 3247 GCTCCCAGCTCCGCAGCCGT 1695 3248 TGCTCCCAGCTCCGCAGCCG 1694 3249 CTGCTCCCAGCTCCGCAGCC 1693 3250 ACTGCTCCCAGCTCCGCAGC 1692 3251 CACTGCTCCCAGCTCCGCAG 1691 3252 CCACTGCTCCCAGCTCCGCA 1690 3253 GCCACTGCTCCCAGCTCCGC 1689 3254 AGCCACTGCTCCCAGCTCCG 1688 3255 CTTCACCGATTGTCTAAACAGAGAC 1522 3256 TTCGTTCCCGGTGGGCTCGAGGGC 35 3257 TCGTTCCCGGTGGGCTCGAG 36 3258 CGTTCCCGGTGGGCTCGAGG 37 3259 GTTCCCGGTGGGCTCGAGGG 38 3260 TTCCCGGTGGGCTCGAGGGC 39 3261 TCCCGGTGGGCTCGAGGGCA 40 3262 CCCGGTGGGCTCGAGGGCAG 41 3263 CCGGTGGGCTCGAGGGCAGA 42 3264 TTTCGTTCCCGGTGGGCTCG 34 3265 CTTTCGTTCCCGGTGGGCTC 33 3266 TCTTTCGTTCCCGGTGGGCT 32 3267 CTCTTTCGTTCCCGGTGGGC 31 3268 TCTCTTTCGTTCCCGGTGGG 30 3269 TTCTCTTTCGTTCCCGGTGG 29 3270 CTTCTCTTTCGTTCCCGGTG 28 3271 GCTTCTCTTTCGTTCCCGGT 27 3272 AGCTTCTCTTTCGTTCCCGG 26 3273 GAGCTTCTCTTTCGTTCCCG 25 3274 AGAGCTTCTCTTTCGTTCCC 24 3275 TAGAGCTTCTCTTTCGTTCC 23 3276 TGCTTCCGCGTCGGCACCCAAG 1150 3277 GCTTCCGCGTCGGCACCCAA 1151 3278 CTTCCGCGTCGGCACCCAAG 1152 3279 TTCCGCGTCGGCACCCAAGA 1153 3280 TCCGCGTCGGCACCCAAGAA 1154 3281 CCGCGTCGGCACCCAAGAAT 1155 3282 CGCGTCGGCACCCAAGAATT 1156 3283 GCGTCGGCACCCAAGAATTT 1157 3284 CGTCGGCACCCAAGAATTTC 1158 3285 GTCGGCACCCAAGAATTTCT 1159 3286 TCGGCACCCAAGAATTTCTT 1160 3287 CGGCACCCAAGAATTTCTTA 1161 3288 CTGCTTCCGCGTCGGCACCC 1149 3289 TCTGCTTCCGCGTCGGCACC 1148 3290 ATCTGCTTCCGCGTCGGCAC 1147 3291 AATCTGCTTCCGCGTCGGCA 1146 3292 GAATCTGCTTCCGCGTCGGC 1145 3293 TGAATCTGCTTCCGCGTCGG 1144 3294 CTGAATCTGCTTCCGCGTCG 1143 3295 TCTGAATCTGCTTCCGCGTC 1142 3296 CTCTGAATCTGCTTCCGCGT 1141 3297 GCTCTGAATCTGCTTCCGCG 1140 3298 GGCTCTGAATCTGCTTCCGC 1139 3299 AGGCTCTGAATCTGCTTCCG 1138

Hot Zones (Relative upstream location to gene start site)  1-800 1050-1250 1400-1800 2850-3400

Examples

In FIG. 31, In MCF7 (human mammary breast cell line), IL6_(—)1 (145) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The IL6 sequence IL6_(—)1 (145) fits the independent and dependent DNAi motif claims.

The secondary structure for IL6_(—)1 (145) is shown in FIG. 32.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11963) AGGGACCTCCCCAGCCATGGGGGCAGGGCCAAATGGGGCTTCTTCA GGACCAGCAAAGCCATTTTTCTCATCAGCAAACTAGCTTCAGAGAAGTTT GCAATCAGGGCACTCTCTTCCAAGCCTAGAGACCCAGGGAAAGGGGTACG GGGGTGTCCCAAGGCAAAGAGAATCTACACTTTTTGCCCCCGGAGAGGCT ACTTCCCTCCCAAGATGCCTGGGATTTTCCACTTCAGCAGGGGGAAGGTA AGTCACATAGCAAAATAATGAGGGCACAGAACAGATGACCTCCCTATAGA GTTTTGAATGAGAAACACAGCAGGGCAGATGTGCCCCTTCTCTAGTCTAG GAGGAGCTAGGTCCAGCCCCTGAACATCCTCCCCCTCAGAAAAGCTGAGG CCAGACTAAGAATTCACCAGACCAAGGAGCTACAACAGGACATCAGAGCT GAGGCTGCAAAGCCAGGACTGAGACCAGACCAGGCAGGAAACTGTCAAGA GCTTTGGTCACCAGGCCTGGCTGCCCTCCAACATCAGCTGGCTCTTTCTA AATTGACACACCACATGTCCCTAAAATTCTCTCTTCAAGTAATACCACCA TCAAAGCAGGACATTTCCCAGAGCCTTAGAGCCTGGTGTCTGCTCAGTGG GACTCAACCCCAGAAGAAGCTGTTAAATCACCCACTGTTTCAGTTTACAA ACTTCTTACGACTTGGCAACAAGTGAAACTACATTCTGGCAGCAACTGCA AGTTCCCTAGTACCCAGGACTTCCCGTTTTTTCTTGCTGTACTCCCTCCT GTTAAATCACAGACTCATCCATCTCCAACCCCCAGAATATAGAGAAAGAG CACAACACTACATCTTAACTCCTGAGACGTGGAGAACACTTCTCCTCCTG AGAGCTTAAGTACCAAATGGAAGCTACTTTTCCCCCTTGGTCTCAAATGT ATTACTAGATTCTGAACTGGACTCCACCATCACGTAAGAAAGCAGTCATG GGCAGTAATTCTGGGAGATCCAGATAGGACATGCCAGCCCCACACTGGTG GCATAGGAAGCCAAGTTGCTGCTTCCTCCCTGTGCACTCCCATTTGTCTG GCCTCTCTTGATCTCAGCTGGCGCTCACTTCACATCAGCTATGATGCAAT CCAGCAACTAAAGTATTAGTTAATAAATGCTGACAGCACAGCCTTTTCTG GTCACGTATTCATACTAAAATACGGGGGAGAGTTGGGGGGAGAGGGGGAT ATATGGGAAATCTCTGTACCTTCCTCTCCATTTTGCTATGACCTAAAGCT GCCCTTTAAAAAATACAAGGGGCTGGGCACAGTGGTTCACGCCTGTAAAC CCAGCACTTTGGGAGGCCGAGGCGCGTGGATCACCTGAGGTCAGGAGTTC AAGACCCGCCTGGCCAACATGGCAAAACCCCGTTTCTACTAAAAATACAA AAAGTAGCTGGGCGTGGTCGCATGCATCTGTAGTCCCAGCTACTCAGGAG GCTGAGGCAAGAGAATTGCTTGAACCTGGGAGGCGGCGGTTGAAGTGAGC CAAGATCATGCCATTGCCCTCCAGCCTGGGCAACAGAGCAAGACTCCTTC TCAAGAGAAAAAACAAAACAAAACAAGAAAAAACAAAGAATGAGCTCTCC ACGCGAAAAATCCATTGAGATGCAAAGGAAGGAAGCTATCATTGTGGAAT TGCACATGTCAGTTACATTAACGTTTTTGGAGCAAGGTAGAGCTCATCTC TCCCACAAGCAAATTCCAGCCCAAAGCATTGATACTAATAAAGTGCCATG CTGCGATGTGCAGGGGGCAGACAGTGTCTCCAAGCTCCCTACACACATGC CTTCCCACAGTTTGCCCTTTCTTGACCCCAGAAGCATCAGGCCCCTTCAC CCTCGAGGGCCACTATCAGGAGTTTGAATTAATGGCAATCACCATGCACA GGGAAGGCTGTGGAATTCTGACATAAAAACACTTAGTGGAGGGCTTGGAA AAAGTCTAGTAGGAGCAAGACGCAAGCTGGACTAATTATCTAAAACAAGA GACCTGGTTTGGGGATCTTAATGTTCTCAAAAAAGAAAATTATTATTATT TTTCATTTTGCACTTTGTGCCATAAAACATTTTCAACAAAACATAGAATC TCATTTCTTTTGAGGGAAAATGATTGGGAGACCAGCTCATTGCTGGCACA GAGGCCTGGTTCATTCATAATTCCTTCATAGGCAAGACACCAGGTGAACC GATATAGCCGAGCTGGAAGAGCTCTCCAAGGCAGAGACTCTGAGCCAAGG AATGTTCAAAGAGCTAGCATGTATTGTGGGATTACTATGCGCCAGGAATT TTTTACACTGCATCACGTTCCATCTTCACAACAGCCCTAGAAAGGAAGAA CTATTATTACCCCCGTTTTATAGGTGAATAAACAAGGGCACAGGTCCTTG ATGTAACAGCCAGGATCAAACAGCTGGGAAGACGAGAAAACCTTTCCCAG GCTAGGATAACAGAGGATTTGGTTGAAAATACAGGCAATTAGGTGCTACC TCTGGGAAAAGGGGCCAGGAGAGGAAGGAGACACTTTTCCCTGCATGCCC TGATGTCCTATTTGAACATTTTATCATGAACACGAACTTCCTATTTAAAA AACACTTTTTATTGAAAAGATAAATCTGTGTGTTGTATTGTGTCACTCAG TTCAAGTACTTGAAATTTATTGAATTGTATTTTCTAAAAAATAGATAGTT GAGTAAAAGCAAGCTCACATTACATAGACGGATCACAGTGCACGGCTGCG GAGCTGGGAGCAGTGGCTTCGTTTCATGCAGGAAAGAGAACTTGGTTCAG GAGTGTCTACGTTGCTTAAGACAGGAGAGCACTAAAAATGAAACCATCCA GCCATCCTCCCCCATTTTCATTTTCACACCAAAGAATCCCACCGCGGCAG AGGACCACCGTCTCTGTTTAGACAATCGGTGAAGAATGGATGACCTCACT TTCCCCAACAGGCGGGTCCTGAAATGTTATGCACGAAACAAAACTTGAGT AAATGCCCAACAGAGGTCACTGTTTTATCGATCTTGAAGAGATCTCTTCT TAGCAAAGCAAAGAAACCGATTGTGAAGGTAACACCATGTTTGGTAAATA AGTGTTTTGGTGTTGTGCAAGGGTCTGGTTTCAGCCTGAAGCCATCTCAG AGCTGTCTGGGTCTCTGGAGACTGGAGGGACAACCTAGTCTAGAGCCCAT TTGCATGAGACCAAGGATCCTCCTGCAAGAGACACCATCCTGAGGGAAGA GGGCTTCTGAACCAGCTTGACCCAATAAGAAATTCTTGGGTGCCGACGCG GAAGCAGATTCAGAGCCTAGAGCCGTGCCTGCGTCCGTAGTTTCCTTCTA GCTTCTTTTGATTTCAAATCAAGACTTACAGGGAGAGGGAGCGATAAACA CAAACTCTGCAAGATGCCACAAGGTCCTCCTTTGACATCCCCAACAAAGA GGTGAGTAGTATTCTCCCCCTTTCTGCCCTGAACCAAGTGGGCTTCAGTA ATTTCAGGGCTCCAGGAGACCTGGGGCCCATGCAGGTGCCCCAGTGAAAC AGTGGTGAAGAGACTCAGTGGCAATGGGGAGAGCACTGGCAGCACAAGGC AAACCTCTGGCACAGAGAGCAAAGTCCTCACTGGGAGGATTCCCAAGGGG TCACTTGGGAGAGGGCAGGGCAGCAGCCAACCTCCTCTAAGTGGGCTGAA GCAGGTGAAGAAAGTGGCAGAAGCCACGCGGTGGCAAAAAGGAGTCACAC ACTCCACCTGGAGACGCCTTGAAGTAACTGCACGAAATTTGAGGATGGCC AGGCAGTTCTACAACAGCCGCTCACAGGGAGAGCCAGAACACAGAAGAAC TCAGATGACTGGTAGTATTACCTTCTTCATAATCCCAGGCTTGGGGGGCT GCGATGGAGTCAGAGGAAACTCAGTTCAGAACATCTTTGGTTTTTACAAA TACAAATTAACTGGAACGCTAAATTCTAGCCTGTTAATCTGGTCACTGAA AAAAAATTTTTTTTTTTTCAAAAAACATAGCTTTAGCTTATTTTTTTTCT CTTTGTAAAACTTCGTGCATGACTTCAGCTTTACTCTTTGTCAAGACATG CCAAAGTGCTGAGTCACTAATAAAAGAAAAAAAGAAAGTAAAGGAAGAGT GGTTCTGCTTCTTAGCGCTAGCCTCAATGACGACCTAAGCTGCACTTTTC CCCCTAGTTGTGTCTTGCCATGCTAAAGGACGTCACATTGCACAATCTTA ATAAGGTTTCCAATCAGCCCCACCCGCTCTGGCCCCACCCTCACCCTCCA ACAAAGATTTATCAAATGTGGGATTTTCCCATGAGTCTCAATATTAGAGT CTCAACCCCCAATAAATATAGGACTGGAGATGTCTGAGGCTCATTCTGCC CTCGAGCCCACCGGGAACGAAAGAGAAGCTCTATCTCCCCTCCAGGAGCC CAGCT ATG

15) IL-23. IL-23 is produced by dendritic cells and macrophages. Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two subunits (p40-S-S-p19): p40, a component of the IL-12 cytokine and p19, the product of the IL23 gene (also considered the IL-23 alpha subunit). IL-23 is an important part of the inflammatory response against infection. Both IL-23 and IL-12 can activate the transcription activator STAT4, and stimulate the production of interferon-gamma (IFNG). In contrast to IL-12, which acts mainly on naive CD4(+) T cells, IL-23 preferentially acts on memory CD4(+) T cells (Oppmann et al., 2001, Immunity 13 (5): 715-25).

IL-23 promotes upregulation of the matrix metalloprotease MMP9, increases angiogenesis and reduces CD8+ T-cell infiltration. In conjunction with IL-6 and TGF-β1, IL-23 stimulates naive CD4+ T cells to differentiate into a novel subset of cells called Th17 cells, which are distinct from the classical Th1 and Th2 cells. Th17 cells produce IL-17, a proinflammatory cytokine that enhances T cell priming and stimulates the production of other proinflammatory molecules such as IL-1, IL-6, TNF-alpha, NOS-2, and chemokines resulting in inflammation.

IL-23 may also play a role in the intestinal immune system which has the challenge of maintaining both a state of tolerance toward intestinal antigens and the ability to combat pathogens. This balance is partially achieved by reciprocal regulation of proinflammatory, effector CD4+ T cells and tolerizing, suppressive regulatory T cells. Inflammatory bowel disease (IBD) comprises Crohn's disease (CD) and ulcerative colitis (UC). Genome-wide association studies have linked CD to a number of IL-23 pathway genes, notably IL23R (interleukin 23 receptor). Similar associations in IL-23 pathway genes have been observed in UC. IL23R is a key differentiation feature of CD4+ Th17 cells, effector cells that are critical in mediating antimicrobial defenses. However, IL-23 and Th17 cell dysregulation can lead to end-organ inflammation. The differentiation of inflammatory Th17 cells and suppressive CD4+ Treg subsets is reciprocally regulated by relative concentrations of TGFβ, with the concomitant presence of proinflammatory cytokines favoring Th17 differentiation. The identification of IL-23 pathway and Th17 expressed genes in IBD pathogenesis highlights the importance of the proper regulation of the IL-23/Th17 pathway in maintaining intestinal immune homeostasis (reviewed in Abraham and Cho, 2009; Ann. Rev. Med. 60: 97-110).

Protein: IL23 Gene: IL23A (Homo sapiens, chromosome 12, 56732663-56734194 [NCBI Reference Sequence: NC_(—)000012.11]; start site location: 56732829; strand: positive)

Gene Identification GeneID 51561 HGNC 15488 HPRD 12026 MIM 605580

Targeted Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 3300 TCCCTGCATTGTAAGGCCCGCC 195 3319 CACAGCGGGGATGGGGTGGGAGGG 414 3320 GACGTCAGAATGAGGCCATCG 1296 3341 GAGCCAGCACGGTGGTGGGCGCC 1651 3365 GCGTTTGTCCCACCGGCGCCCCG 4861 3479 TAACGCCACCCAACAAGTCCGGCG 4830

Target Shift Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 3300 TCCCTGCATTGTAAGGCCCGCC 195 3301 CCCTGCATTGTAAGGCCCGC 196 3302 CCTGCATTGTAAGGCCCGCC 197 3303 CTGCATTGTAAGGCCCGCCC 198 3304 TGCATTGTAAGGCCCGCCCT 199 3305 GCATTGTAAGGCCCGCCCTT 200 3306 CATTGTAAGGCCCGCCCTTT 201 3307 ATTGTAAGGCCCGCCCTTTA 202 3308 TTGTAAGGCCCGCCCTTTAT 203 3309 TGTAAGGCCCGCCCTTTATA 204 3310 GTAAGGCCCGCCCTTTATAC 205 3311 TAAGGCCCGCCCTTTATACC 206 3312 AAGGCCCGCCCTTTATACCA 207 3313 AGGCCCGCCCTTTATACCAG 208 3314 GGCCCGCCCTTTATACCAGC 209 3315 GCCCGCCCTTTATACCAGCA 210 3316 CCCGCCCTTTATACCAGCAG 211 3317 CCGCCCTTTATACCAGCAGG 212 3318 CGCCCTTTATACCAGCAGGT 213 3319 CACAGCGGGGATGGGGTGGGAGGG 414 3320 GACGTCAGAATGAGGCCATCG 1296 3321 ACGTCAGAATGAGGCCATCG 1297 3322 CGTCAGAATGAGGCCATCGG 1298 3323 GTCAGAATGAGGCCATCGGT 1299 3324 TCAGAATGAGGCCATCGGTG 1300 3325 CAGAATGAGGCCATCGGTGA 1301 3326 AGAATGAGGCCATCGGTGAC 1302 3327 GAATGAGGCCATCGGTGACC 1303 3328 AATGAGGCCATCGGTGACCA 1304 3329 ATGAGGCCATCGGTGACCAC 1305 3330 TGAGGCCATCGGTGACCACA 1306 3331 GAGGCCATCGGTGACCACAC 1307 3332 AGGCCATCGGTGACCACACA 1308 3333 GGCCATCGGTGACCACACAG 1309 3334 GCCATCGGTGACCACACAGC 1310 3335 CCATCGGTGACCACACAGCT 1311 3336 CATCGGTGACCACACAGCTG 1312 3337 ATCGGTGACCACACAGCTGG 1313 3338 TCGGTGACCACACAGCTGGC 1314 3339 CGGTGACCACACAGCTGGCT 1315 3340 AGACGTCAGAATGAGGCCAT 1295 3341 GAGCCAGCACGGTGGTGGGCGCC 1651 3342 AGCCAGCACGGTGGTGGGCG 1652 3343 GCCAGCACGGTGGTGGGCGC 1653 3344 CCAGCACGGTGGTGGGCGCC 1654 3345 CAGCACGGTGGTGGGCGCCT 1655 3346 AGCACGGTGGTGGGCGCCTA 1656 3347 GCACGGTGGTGGGCGCCTAT 1657 3348 CACGGTGGTGGGCGCCTATA 1658 3349 ACGGTGGTGGGCGCCTATAG 1659 3350 CGGTGGTGGGCGCCTATAGT 1660 3351 GGTGGTGGGCGCCTATAGTC 1661 3352 GTGGTGGGCGCCTATAGTCC 1662 3353 TGGTGGGCGCCTATAGTCCC 1663 3354 GGTGGGCGCCTATAGTCCCA 1664 3355 GTGGGCGCCTATAGTCCCAG 1665 3356 TGGGCGCCTATAGTCCCAGC 1666 3357 GGGCGCCTATAGTCCCAGCT 1667 3358 GGCGCCTATAGTCCCAGCTA 1668 3359 GCGCCTATAGTCCCAGCTAC 1669 3360 CGCCTATAGTCCCAGCTACT 1670 3361 TGAGCCAGCACGGTGGTGGG 1650 3362 ATGAGCCAGCACGGTGGTGG 1649 3363 AATGAGCCAGCACGGTGGTG 1648 3364 AAATGAGCCAGCACGGTGGT 1647 3365 GCGTTTGTCCCACCGGCGCCCCG 4861 3366 CGTTTGTCCCACCGGCGCCC 4862 3367 GTTTGTCCCACCGGCGCCCC 4863 3368 TTTGTCCCACCGGCGCCCCG 4864 3369 TTGTCCCACCGGCGCCCCGT 4865 3370 TGTCCCACCGGCGCCCCGTA 4866 3371 GTCCCACCGGCGCCCCGTAA 4867 3372 TCCCACCGGCGCCCCGTAAC 4868 3373 CCCACCGGCGCCCCGTAACC 4869 3374 CCACCGGCGCCCCGTAACCT 4870 3375 CACCGGCGCCCCGTAACCTC 4871 3376 ACCGGCGCCCCGTAACCTCT 4872 3377 CCGGCGCCCCGTAACCTCTT 4873 3378 CGGCGCCCCGTAACCTCTTT 4874 3379 GGCGCCCCGTAACCTCTTTT 4875 3380 GCGCCCCGTAACCTCTTTTT 4876 3381 CGCCCCGTAACCTCTTTTTC 4877 3382 GCCCCGTAACCTCTTTTTCC 4878 3383 CCCCGTAACCTCTTTTTCCG 4879 3384 CCCGTAACCTCTTTTTCCGG 4880 3385 CCGTAACCTCTTTTTCCGGC 4881 3386 CGTAACCTCTTTTTCCGGCG 4882 3387 GTAACCTCTTTTTCCGGCGC 4883 3388 TAACCTCTTTTTCCGGCGCG 4884 3389 AACCTCTTTTTCCGGCGCGT 4885 3390 ACCTCTTTTTCCGGCGCGTG 4886 3391 CCTCTTTTTCCGGCGCGTGC 4887 3392 CTCTTTTTCCGGCGCGTGCG 4888 3393 TCTTTTTCCGGCGCGTGCGT 4889 3394 CTTTTTCCGGCGCGTGCGTC 4890 3395 TTTTTCCGGCGCGTGCGTCA 4891 3396 TTTTCCGGCGCGTGCGTCAC 4892 3397 TTTCCGGCGCGTGCGTCACA 4893 3398 TTCCGGCGCGTGCGTCACAC 4894 3399 TCCGGCGCGTGCGTCACACG 4895 3400 CCGGCGCGTGCGTCACACGC 4896 3401 CGGCGCGTGCGTCACACGCT 4897 3402 GGCGCGTGCGTCACACGCTC 4898 3403 GCGCGTGCGTCACACGCTCT 4899 3404 CGCGTGCGTCACACGCTCTC 4900 3405 GCGTGCGTCACACGCTCTCT 4901 3406 CGTGCGTCACACGCTCTCTC 4902 3407 GTGCGTCACACGCTCTCTCC 4903 3408 TGCGTCACACGCTCTCTCCT 4904 3409 GCGTCACACGCTCTCTCCTG 4905 3410 CGTCACACGCTCTCTCCTGG 4906 3411 GTCACACGCTCTCTCCTGGG 4907 3412 TCACACGCTCTCTCCTGGGG 4908 3413 CACACGCTCTCTCCTGGGGT 4909 3414 ACACGCTCTCTCCTGGGGTC 4910 3415 CACGCTCTCTCCTGGGGTCG 4911 3416 ACGCTCTCTCCTGGGGTCGC 4912 3417 CGCTCTCTCCTGGGGTCGCC 4913 3418 GCTCTCTCCTGGGGTCGCCG 4914 3419 CTCTCTCCTGGGGTCGCCGT 4915 3420 TCTCTCCTGGGGTCGCCGTA 4916 3421 CTCTCCTGGGGTCGCCGTAC 4917 3422 TCTCCTGGGGTCGCCGTACC 4918 3423 CTCCTGGGGTCGCCGTACCT 4919 3424 TCCTGGGGTCGCCGTACCTG 4920 3425 CCTGGGGTCGCCGTACCTGG 4921 3426 CTGGGGTCGCCGTACCTGGC 4922 3427 TGGGGTCGCCGTACCTGGCT 4923 3428 GGGGTCGCCGTACCTGGCTC 4924 3429 GGGTCGCCGTACCTGGCTCC 4925 3430 GGTCGCCGTACCTGGCTCCT 4926 3431 GTCGCCGTACCTGGCTCCTT 4927 3432 TCGCCGTACCTGGCTCCTTC 4928 3433 CGCCGTACCTGGCTCCTTCT 4929 3434 GCCGTACCTGGCTCCTTCTG 4930 3435 CCGTACCTGGCTCCTTCTGA 4931 3436 CGTACCTGGCTCCTTCTGAT 4932 3437 TGCGTTTGTCCCACCGGCGC 4860 3438 CTGCGTTTGTCCCACCGGCG 4859 3439 GCTGCGTTTGTCCCACCGGC 4858 3440 GGCTGCGTTTGTCCCACCGG 4857 3441 TGGCTGCGTTTGTCCCACCG 4856 3442 CTGGCTGCGTTTGTCCCACC 4855 3443 TCTGGCTGCGTTTGTCCCAC 4854 3444 GTCTGGCTGCGTTTGTCCCA 4853 3445 CGTCTGGCTGCGTTTGTCCC 4852 3446 GCGTCTGGCTGCGTTTGTCC 4851 3447 GGCGTCTGGCTGCGTTTGTC 4850 3448 CGGCGTCTGGCTGCGTTTGT 4849 3449 CCGGCGTCTGGCTGCGTTTG 4848 3450 TCCGGCGTCTGGCTGCGTTT 4847 3451 GTCCGGCGTCTGGCTGCGTT 4846 3452 AGTCCGGCGTCTGGCTGCGT 4845 3453 AAGTCCGGCGTCTGGCTGCG 4844 3454 CAAGTCCGGCGTCTGGCTGC 4843 3455 ACAAGTCCGGCGTCTGGCTG 4842 3456 AACAAGTCCGGCGTCTGGCT 4841 3457 CAACAAGTCCGGCGTCTGGC 4840 3458 CCAACAAGTCCGGCGTCTGG 4839 3459 CCCAACAAGTCCGGCGTCTG 4838 3460 ACCCAACAAGTCCGGCGTCT 4837 3461 CACCCAACAAGTCCGGCGTC 4836 3462 CCACCCAACAAGTCCGGCGT 4835 3463 GCCACCCAACAAGTCCGGCG 4834 3464 CGCCACCCAACAAGTCCGGC 4833 3465 ACGCCACCCAACAAGTCCGG 4832 3466 AACGCCACCCAACAAGTCCG 4831 3467 TAACGCCACCCAACAAGTCC 4830 3468 CTAACGCCACCCAACAAGTC 4829 3469 TCTAACGCCACCCAACAAGT 4828 3470 TTCTAACGCCACCCAACAAG 4827 3471 TTTCTAACGCCACCCAACAA 4826 3472 CTTTCTAACGCCACCCAACA 4825 3473 ACTTTCTAACGCCACCCAAC 4824 3474 TACTTTCTAACGCCACCCAA 4823 3475 TTACTTTCTAACGCCACCCA 4822 3476 GTTACTTTCTAACGCCACCC 4821 3477 AGTTACTTTCTAACGCCACC 4820 3478 GAGTTACTTTCTAACGCCAC 4819 3479 TAACGCCACCCAACAAGTCCGGCG 4830 3480 AACGCCACCCAACAAGTCCG 4831 3481 ACGCCACCCAACAAGTCCGG 4832 3482 CGCCACCCAACAAGTCCGGC 4833 3483 GCCACCCAACAAGTCCGGCG 4834 3484 CCACCCAACAAGTCCGGCGT 4835 3485 CACCCAACAAGTCCGGCGTC 4836 3486 ACCCAACAAGTCCGGCGTCT 4837 3487 CCCAACAAGTCCGGCGTCTG 4838 3488 CCAACAAGTCCGGCGTCTGG 4839 3489 CAACAAGTCCGGCGTCTGGC 4840 3490 AACAAGTCCGGCGTCTGGCT 4841 3491 ACAAGTCCGGCGTCTGGCTG 4842 3492 CAAGTCCGGCGTCTGGCTGC 4843 3493 AAGTCCGGCGTCTGGCTGCG 4844 3494 AGTCCGGCGTCTGGCTGCGT 4845 3495 GTCCGGCGTCTGGCTGCGTT 4846 3496 TCCGGCGTCTGGCTGCGTTT 4847 3497 CCGGCGTCTGGCTGCGTTTG 4848 3498 CGGCGTCTGGCTGCGTTTGT 4849 3499 GGCGTCTGGCTGCGTTTGTC 4850 3500 GCGTCTGGCTGCGTTTGTCC 4851 3501 CGTCTGGCTGCGTTTGTCCC 4852 3502 GTCTGGCTGCGTTTGTCCCA 4853 3503 TCTGGCTGCGTTTGTCCCAC 4854 3504 CTGGCTGCGTTTGTCCCACC 4855 3505 TGGCTGCGTTTGTCCCACCG 4856 3506 GGCTGCGTTTGTCCCACCGG 4857 3507 GCTGCGTTTGTCCCACCGGC 4858 3508 CTGCGTTTGTCCCACCGGCG 4859 3509 TGCGTTTGTCCCACCGGCGC 4860 3510 GCGTTTGTCCCACCGGCGCC 4861 3511 CGTTTGTCCCACCGGCGCCC 4862 3512 GTTTGTCCCACCGGCGCCCC 4863 3513 TTTGTCCCACCGGCGCCCCG 4864 3514 TTGTCCCACCGGCGCCCCGT 4865 3515 TGTCCCACCGGCGCCCCGTA 4866 3516 GTCCCACCGGCGCCCCGTAA 4867 3517 TCCCACCGGCGCCCCGTAAC 4868 3518 CCCACCGGCGCCCCGTAACC 4869 3519 CCACCGGCGCCCCGTAACCT 4870 3520 CACCGGCGCCCCGTAACCTC 4871 3521 ACCGGCGCCCCGTAACCTCT 4872 3522 CCGGCGCCCCGTAACCTCTT 4873 3523 CGGCGCCCCGTAACCTCTTT 4874 3524 GGCGCCCCGTAACCTCTTTT 4875 3525 GCGCCCCGTAACCTCTTTTT 4876 3526 CGCCCCGTAACCTCTTTTTC 4877 3527 GCCCCGTAACCTCTTTTTCC 4878 3528 CCCCGTAACCTCTTTTTCCG 4879 3529 CCCGTAACCTCTTTTTCCGG 4880 3530 CCGTAACCTCTTTTTCCGGC 4881 3531 CGTAACCTCTTTTTCCGGCG 4882 3532 GTAACCTCTTTTTCCGGCGC 4883 3533 TAACCTCTTTTTCCGGCGCG 4884 3534 AACCTCTTTTTCCGGCGCGT 4885 3535 ACCTCTTTTTCCGGCGCGTG 4886 3536 CCTCTTTTTCCGGCGCGTGC 4887 3537 CTCTTTTTCCGGCGCGTGCG 4888 3538 TCTTTTTCCGGCGCGTGCGT 4889 3539 CTTTTTCCGGCGCGTGCGTC 4890 3540 TTTTTCCGGCGCGTGCGTCA 4891 3541 TTTTCCGGCGCGTGCGTCAC 4892 3542 TTTCCGGCGCGTGCGTCACA 4893 3543 TTCCGGCGCGTGCGTCACAC 4894 3544 TCCGGCGCGTGCGTCACACG 4895 3545 CCGGCGCGTGCGTCACACGC 4896 3546 CGGCGCGTGCGTCACACGCT 4897 3547 GGCGCGTGCGTCACACGCTC 4898 3548 GCGCGTGCGTCACACGCTCT 4899 3549 CGCGTGCGTCACACGCTCTC 4900 3550 GCGTGCGTCACACGCTCTCT 4901 3551 CGTGCGTCACACGCTCTCTC 4902 3552 GTGCGTCACACGCTCTCTCC 4903 3553 TGCGTCACACGCTCTCTCCT 4904 3554 GCGTCACACGCTCTCTCCTG 4905 3555 CGTCACACGCTCTCTCCTGG 4906 3556 GTCACACGCTCTCTCCTGGG 4907 3557 TCACACGCTCTCTCCTGGGG 4908 3558 CACACGCTCTCTCCTGGGGT 4909 3559 ACACGCTCTCTCCTGGGGTC 4910 3560 CACGCTCTCTCCTGGGGTCG 4911 3561 ACGCTCTCTCCTGGGGTCGC 4912 3562 CGCTCTCTCCTGGGGTCGCC 4913 3563 GCTCTCTCCTGGGGTCGCCG 4914 3564 CTCTCTCCTGGGGTCGCCGT 4915 3565 TCTCTCCTGGGGTCGCCGTA 4916 3566 CTCTCCTGGGGTCGCCGTAC 4917 3567 TCTCCTGGGGTCGCCGTACC 4918 3568 CTCCTGGGGTCGCCGTACCT 4919 3569 TCCTGGGGTCGCCGTACCTG 4920 3570 CCTGGGGTCGCCGTACCTGG 4921 3571 CTGGGGTCGCCGTACCTGGC 4922 3572 TGGGGTCGCCGTACCTGGCT 4923 3573 GGGGTCGCCGTACCTGGCTC 4924 3574 GGGTCGCCGTACCTGGCTCC 4925 3575 GGTCGCCGTACCTGGCTCCT 4926 3576 GTCGCCGTACCTGGCTCCTT 4927 3577 TCGCCGTACCTGGCTCCTTC 4928 3578 CGCCGTACCTGGCTCCTTCT 4929 3579 GCCGTACCTGGCTCCTTCTG 4930 3580 CCGTACCTGGCTCCTTCTGA 4931 3581 CGTACCTGGCTCCTTCTGAT 4932 3582 CTAACGCCACCCAACAAGTC 4829 3583 TCTAACGCCACCCAACAAGT 4828 3584 TTCTAACGCCACCCAACAAG 4827 3585 TTTCTAACGCCACCCAACAA 4826 3586 CTTTCTAACGCCACCCAACA 4825 3587 ACTTTCTAACGCCACCCAAC 4824 3588 TACTTTCTAACGCCACCCAA 4823 3589 TTACTTTCTAACGCCACCCA 4822 3590 GTTACTTTCTAACGCCACCC 4821 3591 AGTTACTTTCTAACGCCACC 4820 3592 GAGTTACTTTCTAACGCCAC 4819

Hot Zones (Relative upstream location to gene start site)  1-500  950-1400 1450-1800 3390-4050 4300-5000

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11964) AACTCCCATCCGTGATTGTTCCCTCCCCAGAGACCCCGGTAACATTCCCG GGTAACAAGATGCCCCTGGTTATCAAATTCCCCTAGCTCTTGAGGCTGGC TGGACGTTATCCCTCAGAGGGGGATGAGCATGGCAAATTGGGACTTGTTA TTCTGAAGGATTCGTGGGTCCTGTGAACTCTAATTACTTTGAAATGGGTC TAGGTTGTGAGATGTCTCAGAGCACTTTAGCTCAGCTGTTATTACTGTTT CTAAAGGCCACATAAAGGGACTCTGATGGGAGACATTCCTCATGGAGGAT TCAATTCTATAACATTTCTCTCAATAAAGGCTGGTAAATAGACCTTCATT AAAGGAACCAAGAATTTAAATTTCTAGGACTCAGAGGGGTGGGGTCCTAT ACCCAGTCAGAGATCCTACCTAGAGCCTAGACCAAGAGAAAAACACAGAT GGTCTCTCAAACTGATTTGATCTGACTTCGCAGGTCATTAGATATAGAAT CTCCGAAAAAGGTGGATGCTGAGAGACATAGACAGTTCCTACACTTTAAG AAATCTCCATCTTGAGGTCTCAAATTGAGAAAGACTTAACAGACCCATGA GAGTTACAGATCCCTAATAACCTGGGCTAAATAATCCATGTCTGCCGGGC GCAGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGTGGGCG GATCACCTGAGGTCGGGAGTTCGAGACCAACCTGACCAATATGGAGAAAC CCCGTCTCCACTAAAAATACAAAATTAGCCGGGCCTGGTGACGGGAGCCT GTAATCCCAGCTACTTGGGAGGCTGAGGCAGGAGAATCATTTGAACCTGG GAGGCGGAGGTTGCAGTGAGCCGAGATGGCACCATTGCACTCCAGCCTGG TCAGTAAGAGCGAAACTCCGTCTCAAAAAAAAGAAAAAAAAAAAGAAAAG AAAAAAGGATACTGTGAGGAGACACAAGAGCATCCATGACATAGATTATT TAGCTCAGCTGTAATTACTGTTTCTAATACAGTAATATTAGATGGTGATC TGCCTGCCTCGGCCTCCCAAAGTTCTGGGATTACAGGTGTGAGCCACCGC GCCCAGCCTTTTTTTTTTTTTTTTTTTGAGACAGGATCTCACTCTGTTGC CCATGCTTAAGCGCATTGGCCCTCTCACTCACTGTAGCCTCAACCTCCTG GGCTCAAGCGATCCTCCCACTTCAGCCTCCCAACTAGCTGTAACTACAGG CACTGGCCACCAAACCCAGATAATTTTTTTTTTCCTGTAGAGGTGGGGTT TTGCCACGTTACCCAGGCTGGTCTTGAACTCTTAAGCTCAAGCGATCCTC CTGCCTCGGCCCCCCAAAGTTCTGGGATTACAGGCATGAGCCACCATACC TGGCGTACAGTATCCAGTGTAATGCAGTGATTAAAAATTCAGGATCCAGA CCGGGATGGTGGCTTGTGCCTGTAGTCCCAGGGGTGGAGGTTGCAGTGAA CGGAAATGGTGCCACTGCATTCCAGCCTGGGTGACAGAGTGAGACCCTGT CTCAACAAAACCCCCCAAAAACCAAGAACAAAAAAGAATGCAGGATCTGA TGCTAGATTGTCTGCATTAGAACTCTAGCCACTTAAGCTGGGTGTGGTGG CTCATGCCTGTAATCCCAGCACTTTTGGAGGCCGCAGGCGGGTGGATCAC CTGAGATTGGGAGTTCAAGACCAGCCTGACCAACATGGAGAAACCCTGTT TCTACTAAAAATACAAAATTAGCCACCGGGCATGGTGGTGCATGCCTGTA ATCCCAGCTACTTGGGAGGCTGAGGCAGGAGAATCACTTGAACCCGGGAG GCAGAGGTTACGGTGAGATGAGATGGCACCATTGCACTCCAGCCTGGGCA ACAAGAGCGAAACTCCATCTCAAAAAAAAAAAAAAAAAAAAAGGAATTCT AGCCACTTAGAAGCTCTGTGATCTTGGGCAAATTGCTTATCTTTGCACCT CAGCCTCCTCCTCTGTAATATAGGGTAATAGTATCTACCTTAAAGGGTTG TTGTGAAAATTAAATAGTTTAGTACATGTAAAGTGCTTAGACAAAGTATT TGGCATTAAGCGAGAGTTGGATATATTAGCCATCATTATTAACCACCTGG GGGAACTTCAACTGATTTGGAGTCTAGGCATACAACTGGAAAGACCTGCC TAGGAGTGTCTTGTGAATGCGATTTGCATAACGGTTTAGGCCCAGCTGAC GTCAAGGGCTCCTTATAGCTCCAGGTCAGTTGTAGCCCTGGATGTAGTTC CTGCCACGCAACAGTCCCACAATCTCCCCACCAACCCTTCTTCCTACCCA ACTCCTGCAGCACCAGGAAGTGAAACAAAGAGGCAGAGCCCTGTGCCTCC AACTCACCCTTGTCCCTCTCATCCCATCCCCCAGGCTCTACTTCCTCCTC CTTTTCATCTTTCTTTCATCTCTTATCTTTTAGGGCTCCCAGAATGGGGA CCAGAGATGGGAAGAACATAGGAGACGTTGTACACAAGTAAGGTGAACTC CCTATCCTGCCCCCTCCCCTTTCCTTATTCCATTGGTGTCCACCTTATTA GGGAGAGAGGCAAAACAGTTCTCACCCAAACTCAGATAATTCTCTGATGC TGGAAATGTTTAATCTAAAGGGTAGATTTCCATTTTTTTTTTTTTTTTTT TGAGACAGAGTCTTGCTCTGTCACCCAGGCTGGAGTGCAGTGGCGCCATC TCGGCTCACTGCAACCTCTGCCTCCTGGCTTCAAGCGATTCTCTTGCCTC AGCCTCCCGAGTAGCTGGGACTATAGGCGCCCACCACCGTGCTGGCTCAT TTTTGTATTTTTAGTAGAGACAGGATTTCACCATGATGGCCAGGCTGGTC TCGAACTCCTGACCTCATGATCCGCCTGCCTCGGCCTCCCAAAGTACTGG GATTACAGGCGTGAGCCACTGTACCCGGCCCTTGGTAGATTTAACTTAGA ATCGTAATATTTTTTTTTCTTCTCTTAGCTCATACCTACAGAATCATAAT ATTTGAACCAGAAGTGTCATTGGGCAGTTTTGAATAGCTCTAAGGGAAGG GAGACCTCCATTCAGGACAAGTTTCTCAGAAGAAAAGGGTCAACCTCTTG GGGGAGGCTTTGGGAGCCAGCTGTGTGGTCACCGATGGCCTCATTCTGAC GTCTTCGAAATTGTTCTGGGACCCTCCACTGGGGTCGGGGCAGTCCCGGC TTTGGACCACCTTCCACTCCCACGCCCAACCTCACACTCTTAGCTGTTTC ACTCGATGTTGCATCATGGAGGGTGATGAAATCGGTGTCAGTGGATTTTA CCCATGGATGCAACAAGCTGAAGGACCAGCCAGAGTCATTGACAGTGCAC CTTCGACTACCCAGAACTCCTGGGCTTCCTAGCCATGGGGTCCAAAGCTG GGACTGCCCCGACCCCAGTGGAGGGTCCCAGAACAATTTGGATGACGTCA GAATGAGGCCATGGGACTAGGTGCTGGAATGTCTAAGTTGAACTTCCAGG CCTTATTTGCACTAGTCCTGAAAAAAACATCATCCAACTCTTATAGAGCC TATGAAATCTTGGGCCACTAGGGTTGAGGAGTCAGGTGGTTCTTAGTCAA TAACCCTCTTCCCACAAGAGCCTTTCTAACCTCCACTGTGAGGCCTGAAA TGGGGAGCAATAAGACCTCATACTGGCTTCCCAGTTCTCCAAGTTCCTTC ATGCGCATTCTCTCCCATGAAACCAGGACCATCCAGTTGAAATAATGTTG TTTCCAACTGAGAAAAAGAAGCCCGTTTATTCCTAATAGGGGGCATCAGG TAGGAATCAAACTTCATTGCAAACAGCTCACCATCCTATTGGGAGATGAA TGGATGTTTCTCTGTTTTGCTTTTTCCTCAAGCAGGAGGAAGTGAGGAAA TTAGGTTTGGGGTGGGGTAGGGGTATAGCTTTGAGAGGCAAAAAGATCAG GGAAAGATCAAACAGGAAGGAACTTGAGACCAGATTAATTTAAATATTTG TTCTCCCTTACCCCTCCCACCCCATCCCCGCTGTGCCCCCCATCCCCGCC CCTTCTATAGCTATTTCGATTCCTGGAGAGCATTACACATGTGTCCCATC CCAGGCCTCTAGCCACAGCAACCACACTACTCATTTCCCCTGGAACTGAG GCTGCATACCTGGGCTCCCCACAGAGGGGGATGATGCAGGGAGGGGAATC CCACCTGCTGTGAGTCACCTGCTGGTATAAAGGGCGGGCCTTACAATGCA GGGACCTTAAAAGACTCAGAGACAAAGGGAGAAAAACAACAGGAAGCAGC TTACAAACTCGGTGAACAACTGAGGGAACCAAACCAGAGACGCGCTGAAC AGAGAGAATCAGGCTCAAAGCAAGTGGAAGTGGGCAGAGATTCCACCAGG ACTGGTGCAAGGCGCAGAGCCAGCCAGATTTGAGAAGAAGGCAAAAAG ATG

16) AKT. Akt (Protein kinase B, PKB) is a serine/threonine kinase is an important node several signaling cascades downstream of growth factor receptor tyrosine kinases. Akt plays an essential role in cell survival and altered activity has been associated with cancer and other disease conditions, such as diabetes mellitus, neurodegenerative diseases, and muscle hypotrophy. AKT plays a key role in regulating tumor formation, cell survival, insulin signaling and metabolism (lipid and glucose), growth, migration, proliferation, polarity, cell cycle progression, muscle and cardiomyocyte contractility, angiogenesis, and self-renewal of stem cells (reviewed by Liao and Hung, Am J Transl Res. 2010; 2(1): 19-42). Akt is a downstream mediator of the PI 3-K pathway, resulting in the recruitment of Akt to the plasma membrane via the PH (plexstrin homology domain) of Akt. Akt is fully activated by phosphorylation at two key sites: Ser308 (phosphorylated by PDK1) and Thr478 (phosphorylated by mTOR and DNA-PK). Akt can then phosphorylated a wide range of substrates including transcription factors (e.g. FOXO1), kinases (GSK-3, Raf-1, ASK, Chk1) and other proteins with important signaling roles (e.g. Bad, MDM2).

Protein: AKT1 Gene: AKT1 (Homo sapiens, chromosome 14, 105235686-105262080 [NCBI Reference Sequence: NC_(—)000014.8]; start site location: 105258980; strand: negative)

Gene Identification GeneID 207 HGNC 391 HPRD 01261 MIM 164730

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 3593 GAGGCTCCCGCGACGCTCACGCG 8 3646 TACCGGGCGTCTCAGGTTTTGCC 843 3669 TCCGAGCCGCGCACGCCTCAGGC 1562 3703 CACCAACGGACTCCGTCCGCCC 2010 3770 CCGCCGGCTGCCTCGCTGGCCCAGCG 2464 3927 TCTCGGGTCCCGGCCTCGCCCGGCGG 2556 AGC 4084 CATTCTGGCGGCGCCGCGGCTCGCG 2730 4228 CACCGGGCCGCCGCGTCCGGGCGCG 2838 4338 AKT4 CACATCCGCCTCCGCCGCCCGG 3160

Targeted Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 3593 GAGGCTCCCGCGACGCTCACGCG 8 3594 AGGCTCCCGCGACGCTCACG 9 3595 GGCTCCCGCGACGCTCACGC 10 3596 GCTCCCGCGACGCTCACGCG 11 3597 CTCCCGCGACGCTCACGCGC 12 3598 TCCCGCGACGCTCACGCGCT 13 3599 CCCGCGACGCTCACGCGCTC 14 3600 CCGCGACGCTCACGCGCTCC 15 3601 CGCGACGCTCACGCGCTCCT 16 3602 GCGACGCTCACGCGCTCCTC 17 3603 CGACGCTCACGCGCTCCTCT 18 3604 GACGCTCACGCGCTCCTCTC 19 3605 ACGCTCACGCGCTCCTCTCA 20 3606 CGCTCACGCGCTCCTCTCAG 21 3607 GCTCACGCGCTCCTCTCAGG 22 3608 CTCACGCGCTCCTCTCAGGC 23 3609 TCACGCGCTCCTCTCAGGCT 24 3610 CACGCGCTCCTCTCAGGCTG 25 3611 ACGCGCTCCTCTCAGGCTGG 26 3612 CGCGCTCCTCTCAGGCTGGC 27 3613 GCGCTCCTCTCAGGCTGGCG 28 3614 CGCTCCTCTCAGGCTGGCGC 29 3615 GCTCCTCTCAGGCTGGCGCT 30 3616 CTCCTCTCAGGCTGGCGCTC 31 3617 TCCTCTCAGGCTGGCGCTCC 32 3618 CCTCTCAGGCTGGCGCTCCC 33 3619 CTCTCAGGCTGGCGCTCCCC 34 3620 TCTCAGGCTGGCGCTCCCCG 35 3621 CTCAGGCTGGCGCTCCCCGA 36 3622 TCAGGCTGGCGCTCCCCGAG 37 3623 CAGGCTGGCGCTCCCCGAGC 38 3624 AGGCTGGCGCTCCCCGAGCC 39 3625 GGCTGGCGCTCCCCGAGCCC 40 3626 GCTGGCGCTCCCCGAGCCCA 41 3627 CTGGCGCTCCCCGAGCCCAG 42 3628 TGGCGCTCCCCGAGCCCAGC 43 3629 GGCGCTCCCCGAGCCCAGCT 44 3630 GCGCTCCCCGAGCCCAGCTG 45 3631 CGCTCCCCGAGCCCAGCTGG 46 3632 GCTCCCCGAGCCCAGCTGGC 47 3633 CTCCCCGAGCCCAGCTGGCC 48 3634 TCCCCGAGCCCAGCTGGCCT 49 3635 CCCCGAGCCCAGCTGGCCTG 50 3636 CCCGAGCCCAGCTGGCCTGG 51 3637 CCGAGCCCAGCTGGCCTGGC 52 3638 CGAGCCCAGCTGGCCTGGCC 53 3639 CGAGGCTCCCGCGACGCTCA 7 3640 CCGAGGCTCCCGCGACGCTC 6 3641 CCCGAGGCTCCCGCGACGCT 5 3642 GCCCGAGGCTCCCGCGACGC 4 3643 TGCCCGAGGCTCCCGCGACG 3 3644 GTGCCCGAGGCTCCCGCGAC 2 3645 GGTGCCCGAGGCTCCCGCGA 1 3646 TACCGGGCGTCTCAGGTTTTGCC 843 3647 ACCGGGCGTCTCAGGTTTTG 844 3648 CCGGGCGTCTCAGGTTTTGC 845 3649 CGGGCGTCTCAGGTTTTGCC 846 3650 GGGCGTCTCAGGTTTTGCCA 847 3651 GGCGTCTCAGGTTTTGCCAG 848 3652 GCGTCTCAGGTTTTGCCAGG 849 3653 CGTCTCAGGTTTTGCCAGGC 850 3654 GTACCGGGCGTCTCAGGTTT 842 3655 TGTACCGGGCGTCTCAGGTT 841 3656 ATGTACCGGGCGTCTCAGGT 840 3657 CATGTACCGGGCGTCTCAGG 839 3658 ACATGTACCGGGCGTCTCAG 838 3659 AACATGTACCGGGCGTCTCA 837 3660 CAACATGTACCGGGCGTCTC 836 3661 CCAACATGTACCGGGCGTCT 835 3662 GCCAACATGTACCGGGCGTC 834 3663 GGCCAACATGTACCGGGCGT 833 3664 TGGCCAACATGTACCGGGCG 832 3665 TTGGCCAACATGTACCGGGC 831 3666 TTTGGCCAACATGTACCGGG 830 3667 ATTTGGCCAACATGTACCGG 829 3668 CATTTGGCCAACATGTACCG 828 3669 TCCGAGCCGCGCACGCCTCAGGC 1562 3670 CCGAGCCGCGCACGCCTCAG 1563 3671 CGAGCCGCGCACGCCTCAGG 1564 3672 GAGCCGCGCACGCCTCAGGC 1565 3673 AGCCGCGCACGCCTCAGGCA 1566 3674 GCCGCGCACGCCTCAGGCAC 1567 3675 CCGCGCACGCCTCAGGCACA 1568 3676 CGCGCACGCCTCAGGCACAG 1569 3677 GCGCACGCCTCAGGCACAGG 1570 3678 CGCACGCCTCAGGCACAGGG 1571 3679 GCACGCCTCAGGCACAGGGG 1572 3680 CACGCCTCAGGCACAGGGGG 1573 3681 ACGCCTCAGGCACAGGGGGC 1574 3682 CGCCTCAGGCACAGGGGGCT 1575 3683 CTCCGAGCCGCGCACGCCTC 1561 3684 GCTCCGAGCCGCGCACGCCT 1560 3685 GGCTCCGAGCCGCGCACGCC 1559 3686 GGGCTCCGAGCCGCGCACGC 1558 3687 AGGGCTCCGAGCCGCGCACG 1557 3688 CAGGGCTCCGAGCCGCGCAC 1556 3689 GCAGGGCTCCGAGCCGCGCA 1555 3690 GGCAGGGCTCCGAGCCGCGC 1554 3691 GGGCAGGGCTCCGAGCCGCG 1553 3692 AGGGCAGGGCTCCGAGCCGC 1552 3693 GAGGGCAGGGCTCCGAGCCG 1551 3694 CGAGGGCAGGGCTCCGAGCC 1550 3695 CCGAGGGCAGGGCTCCGAGC 1549 3696 TCCGAGGGCAGGGCTCCGAG 1548 3697 CTCCGAGGGCAGGGCTCCGA 1547 3698 ACTCCGAGGGCAGGGCTCCG 1546 3699 GACTCCGAGGGCAGGGCTCC 1545 3700 GGACTCCGAGGGCAGGGCTC 1544 3701 AGGACTCCGAGGGCAGGGCT 1543 3702 CAGGACTCCGAGGGCAGGGC 1542 3703 CACCAACGGACTCCGTCCGCCC 2010 3704 ACCAACGGACTCCGTCCGCC 2011 3705 CCAACGGACTCCGTCCGCCC 2012 3706 CAACGGACTCCGTCCGCCCT 2013 3707 AACGGACTCCGTCCGCCCTT 2014 3708 ACGGACTCCGTCCGCCCTTC 2015 3709 CGGACTCCGTCCGCCCTTCG 2016 3710 GGACTCCGTCCGCCCTTCGC 2017 3711 GACTCCGTCCGCCCTTCGCT 2018 3712 ACTCCGTCCGCCCTTCGCTC 2019 3713 CTCCGTCCGCCCTTCGCTCG 2020 3714 TCCGTCCGCCCTTCGCTCGG 2021 3715 CCGTCCGCCCTTCGCTCGGA 2022 3716 CGTCCGCCCTTCGCTCGGAT 2023 3717 GTCCGCCCTTCGCTCGGATG 2024 3718 TCCGCCCTTCGCTCGGATGA 2025 3719 CCGCCCTTCGCTCGGATGAG 2026 3720 CGCCCTTCGCTCGGATGAGG 2027 3721 GCCCTTCGCTCGGATGAGGG 2028 3722 CCCTTCGCTCGGATGAGGGA 2029 3723 CCTTCGCTCGGATGAGGGAC 2030 3724 CTTCGCTCGGATGAGGGACT 2031 3725 TTCGCTCGGATGAGGGACTC 2032 3726 TCGCTCGGATGAGGGACTCA 2033 3727 CGCTCGGATGAGGGACTCAA 2034 3728 GCTCGGATGAGGGACTCAAA 2035 3729 CTCGGATGAGGGACTCAAAG 2036 3730 TCGGATGAGGGACTCAAAGC 2037 3731 CCACCAACGGACTCCGTCCG 2009 3732 CCCACCAACGGACTCCGTCC 2008 3733 CCCCACCAACGGACTCCGTC 2007 3734 CCCCCACCAACGGACTCCGT 2006 3735 ACCCCCACCAACGGACTCCG 2005 3736 GACCCCCACCAACGGACTCC 2004 3737 GGACCCCCACCAACGGACTC 2003 3738 CGGACCCCCACCAACGGACT 2002 3739 CCGGACCCCCACCAACGGAC 2001 3740 ACCGGACCCCCACCAACGGA 2000 3741 AACCGGACCCCCACCAACGG 1999 3742 CAACCGGACCCCCACCAACG 1998 3743 GCAACCGGACCCCCACCAAC 1997 3744 GGCAACCGGACCCCCACCAA 1996 3745 AGGCAACCGGACCCCCACCA 1995 3746 GAGGCAACCGGACCCCCACC 1994 3747 AGAGGCAACCGGACCCCCAC 1993 3748 GAGAGGCAACCGGACCCCCA 1992 3749 GGAGAGGCAACCGGACCCCC 1991 3750 GGGAGAGGCAACCGGACCCC 1990 3751 CGGGAGAGGCAACCGGACCC 1989 3752 CCGGGAGAGGCAACCGGACC 1988 3753 CCCGGGAGAGGCAACCGGAC 1987 3754 TCCCGGGAGAGGCAACCGGA 1986 3755 CTCCCGGGAGAGGCAACCGG 1985 3756 GCTCCCGGGAGAGGCAACCG 1984 3757 AGCTCCCGGGAGAGGCAACC 1983 3758 CAGCTCCCGGGAGAGGCAAC 1982 3759 ACAGCTCCCGGGAGAGGCAA 1981 3760 CACAGCTCCCGGGAGAGGCA 1980 3761 ACACAGCTCCCGGGAGAGGC 1979 3762 TACACAGCTCCCGGGAGAGG 1978 3763 CTACACAGCTCCCGGGAGAG 1977 3764 TCTACACAGCTCCCGGGAGA 1976 3765 GTCTACACAGCTCCCGGGAG 1975 3766 AGTCTACACAGCTCCCGGGA 1974 3767 AAGTCTACACAGCTCCCGGG 1973 3768 GAAGTCTACACAGCTCCCGG 1972 3769 AGAAGTCTACACAGCTCCCG 1971 3770 CCGCCGGCTGCCTCGCTGGCCCAGCG 2464 3771 CGCCGGCTGCCTCGCTGGCC 2465 3772 GCCGGCTGCCTCGCTGGCCC 2466 3773 CCGGCTGCCTCGCTGGCCCA 2467 3774 CGGCTGCCTCGCTGGCCCAG 2468 3775 GGCTGCCTCGCTGGCCCAGC 2469 3776 GCTGCCTCGCTGGCCCAGCG 2470 3777 CTGCCTCGCTGGCCCAGCGC 2471 3778 TGCCTCGCTGGCCCAGCGCC 2472 3779 GCCTCGCTGGCCCAGCGCCC 2473 3780 CCTCGCTGGCCCAGCGCCCG 2474 3781 CTCGCTGGCCCAGCGCCCGG 2475 3782 TCGCTGGCCCAGCGCCCGGG 2476 3783 CGCTGGCCCAGCGCCCGGGG 2477 3784 GCTGGCCCAGCGCCCGGGGA 2478 3785 CTGGCCCAGCGCCCGGGGAG 2479 3786 TGGCCCAGCGCCCGGGGAGC 2480 3787 GGCCCAGCGCCCGGGGAGCC 2481 3788 GCCCAGCGCCCGGGGAGCCC 2482 3789 CCCAGCGCCCGGGGAGCCCC 2483 3790 CCAGCGCCCGGGGAGCCCCA 2484 3791 CAGCGCCCGGGGAGCCCCAC 2485 3792 AGCGCCCGGGGAGCCCCACG 2486 3793 GCGCCCGGGGAGCCCCACGG 2487 3794 CGCCCGGGGAGCCCCACGGC 2488 3795 GCCCGGGGAGCCCCACGGCC 2489 3796 CCCGGGGAGCCCCACGGCCC 2490 3797 CCGGGGAGCCCCACGGCCCG 2491 3798 CGGGGAGCCCCACGGCCCGC 2492 3799 GGGGAGCCCCACGGCCCGCA 2493 3800 GGGAGCCCCACGGCCCGCAG 2494 3801 GGAGCCCCACGGCCCGCAGG 2495 3802 GAGCCCCACGGCCCGCAGGG 2496 3803 AGCCCCACGGCCCGCAGGGG 2497 3804 GCCCCACGGCCCGCAGGGGC 2498 3805 CCCCACGGCCCGCAGGGGCA 2499 3806 CCCACGGCCCGCAGGGGCAC 2500 3807 CCACGGCCCGCAGGGGCACC 2501 3808 CACGGCCCGCAGGGGCACCC 2502 3809 ACGGCCCGCAGGGGCACCCC 2503 3810 CGGCCCGCAGGGGCACCCCG 2504 3811 GGCCCGCAGGGGCACCCCGA 2505 3812 GCCCGCAGGGGCACCCCGAG 2506 3813 CCCGCAGGGGCACCCCGAGC 2507 3814 CCGCAGGGGCACCCCGAGCC 2508 3815 CGCAGGGGCACCCCGAGCCC 2509 3816 GCAGGGGCACCCCGAGCCCC 2510 3817 CAGGGGCACCCCGAGCCCCA 2511 3818 AGGGGCACCCCGAGCCCCAG 2512 3819 GGGGCACCCCGAGCCCCAGC 2513 3820 GGGCACCCCGAGCCCCAGCT 2514 3821 GGCACCCCGAGCCCCAGCTC 2515 3822 GCACCCCGAGCCCCAGCTCC 2516 3823 CACCCCGAGCCCCAGCTCCA 2517 3824 ACCCCGAGCCCCAGCTCCAG 2518 3825 CCCCGAGCCCCAGCTCCAGG 2519 3826 CCCGAGCCCCAGCTCCAGGC 2520 3827 CCGAGCCCCAGCTCCAGGCC 2521 3828 CGAGCCCCAGCTCCAGGCCC 2522 3829 GAGCCCCAGCTCCAGGCCCG 2523 3830 AGCCCCAGCTCCAGGCCCGG 2524 3831 GCCCCAGCTCCAGGCCCGGC 2525 3832 CCCCAGCTCCAGGCCCGGCG 2526 3833 CCCAGCTCCAGGCCCGGCGG 2527 3834 CCAGCTCCAGGCCCGGCGGC 2528 3835 CAGCTCCAGGCCCGGCGGCG 2529 3836 AGCTCCAGGCCCGGCGGCGT 2530 3837 GCTCCAGGCCCGGCGGCGTC 2531 3838 CTCCAGGCCCGGCGGCGTCC 2532 3839 TCCAGGCCCGGCGGCGTCCC 2533 3840 CCAGGCCCGGCGGCGTCCCT 2534 3841 CAGGCCCGGCGGCGTCCCTT 2535 3842 AGGCCCGGCGGCGTCCCTTC 2536 3843 GGCCCGGCGGCGTCCCTTCT 2537 3844 GCCCGGCGGCGTCCCTTCTC 2538 3845 CCCGGCGGCGTCCCTTCTCT 2539 3846 CCGGCGGCGTCCCTTCTCTC 2540 3847 CGGCGGCGTCCCTTCTCTCG 2541 3848 GGCGGCGTCCCTTCTCTCGG 2542 3849 GCGGCGTCCCTTCTCTCGGG 2543 3850 CGGCGTCCCTTCTCTCGGGT 2544 3851 GGCGTCCCTTCTCTCGGGTC 2545 3852 GCGTCCCTTCTCTCGGGTCC 2546 3853 CGTCCCTTCTCTCGGGTCCC 2547 3854 GTCCCTTCTCTCGGGTCCCG 2548 3855 TCCCTTCTCTCGGGTCCCGG 2549 3856 CCCTTCTCTCGGGTCCCGGC 2550 3857 CCTTCTCTCGGGTCCCGGCC 2551 3858 CTTCTCTCGGGTCCCGGCCT 2552 3859 TTCTCTCGGGTCCCGGCCTC 2553 3860 TCTCTCGGGTCCCGGCCTCG 2554 3861 CTCTCGGGTCCCGGCCTCGC 2555 3862 TCTCGGGTCCCGGCCTCGCC 2556 3863 CTCGGGTCCCGGCCTCGCCC 2557 3864 TCGGGTCCCGGCCTCGCCCG 2558 3865 CGGGTCCCGGCCTCGCCCGG 2559 3866 GGGTCCCGGCCTCGCCCGGC 2560 3867 GGTCCCGGCCTCGCCCGGCG 2561 3868 GTCCCGGCCTCGCCCGGCGG 2562 3869 TCCCGGCCTCGCCCGGCGGA 2563 3870 CCCGGCCTCGCCCGGCGGAG 2564 3871 CCGGCCTCGCCCGGCGGAGC 2565 3872 CGGCCTCGCCCGGCGGAGCG 2566 3873 GGCCTCGCCCGGCGGAGCGG 2567 3874 GCCTCGCCCGGCGGAGCGGC 2568 3875 CCTCGCCCGGCGGAGCGGCC 2569 3876 CTCGCCCGGCGGAGCGGCCT 2570 3877 TCGCCCGGCGGAGCGGCCTC 2571 3878 CGCCCGGCGGAGCGGCCTCC 2572 3879 GCCCGGCGGAGCGGCCTCCC 2573 3880 CCCGGCGGAGCGGCCTCCCC 2574 3881 CCGGCGGAGCGGCCTCCCCA 2575 3882 CGGCGGAGCGGCCTCCCCAA 2576 3883 GGCGGAGCGGCCTCCCCAAG 2577 3884 GCGGAGCGGCCTCCCCAAGG 2578 3885 CGGAGCGGCCTCCCCAAGGT 2579 3886 GGAGCGGCCTCCCCAAGGTC 2580 3887 GAGCGGCCTCCCCAAGGTCA 2581 3888 AGCGGCCTCCCCAAGGTCAT 2582 3889 GCGGCCTCCCCAAGGTCATG 2583 3890 CGGCCTCCCCAAGGTCATGA 2584 3891 TCCGCCGGCTGCCTCGCTGG 2463 3892 CTCCGCCGGCTGCCTCGCTG 2462 3893 CCTCCGCCGGCTGCCTCGCT 2461 3894 ACCTCCGCCGGCTGCCTCGC 2460 3895 CACCTCCGCCGGCTGCCTCG 2459 3896 GCACCTCCGCCGGCTGCCTC 2458 3897 GGCACCTCCGCCGGCTGCCT 2457 3898 GGGCACCTCCGCCGGCTGCC 2456 3899 GGGGCACCTCCGCCGGCTGC 2455 3900 CGGGGCACCTCCGCCGGCTG 2454 3901 CCGGGGCACCTCCGCCGGCT 2453 3902 CCCGGGGCACCTCCGCCGGC 2452 3903 CCCCGGGGCACCTCCGCCGG 2451 3904 ACCCCGGGGCACCTCCGCCG 2450 3905 AACCCCGGGGCACCTCCGCC 2449 3906 CAACCCCGGGGCACCTCCGC 2448 3907 CCAACCCCGGGGCACCTCCG 2447 3908 TCCAACCCCGGGGCACCTCC 2446 3909 CTCCAACCCCGGGGCACCTC 2445 3910 TCTCCAACCCCGGGGCACCT 2444 3911 TTCTCCAACCCCGGGGCACC 2443 3912 TTTCTCCAACCCCGGGGCAC 2442 3913 CTTTCTCCAACCCCGGGGCA 2441 3914 TCTTTCTCCAACCCCGGGGC 2440 3915 GTCTTTCTCCAACCCCGGGG 2439 3916 AGTCTTTCTCCAACCCCGGG 2438 3917 GAGTCTTTCTCCAACCCCGG 2437 3918 CGAGTCTTTCTCCAACCCCG 2436 3919 GCGAGTCTTTCTCCAACCCC 2435 3920 GGCGAGTCTTTCTCCAACCC 2434 3921 CGGCGAGTCTTTCTCCAACC 2433 3922 GCGGCGAGTCTTTCTCCAAC 2432 3923 CGCGGCGAGTCTTTCTCCAA 2431 3924 CCGCGGCGAGTCTTTCTCCA 2430 3925 GCCGCGGCGAGTCTTTCTCC 2429 3926 GGCCGCGGCGAGTCTTTCTC 2428 3927 TCTCGGGTCCCGGCCTCGCCCGGCGGAGC 2556 3928 CTCGGGTCCCGGCCTCGCCC 2557 3929 TCGGGTCCCGGCCTCGCCCG 2558 3930 CGGGTCCCGGCCTCGCCCGG 2559 3931 GGGTCCCGGCCTCGCCCGGC 2560 3932 GGTCCCGGCCTCGCCCGGCG 2561 3933 GTCCCGGCCTCGCCCGGCGG 2562 3934 TCCCGGCCTCGCCCGGCGGA 2563 3935 CCCGGCCTCGCCCGGCGGAG 2564 3936 CCGGCCTCGCCCGGCGGAGC 2565 3937 CGGCCTCGCCCGGCGGAGCG 2566 3938 GGCCTCGCCCGGCGGAGCGG 2567 3939 GCCTCGCCCGGCGGAGCGGC 2568 3940 CCTCGCCCGGCGGAGCGGCC 2569 3941 CTCGCCCGGCGGAGCGGCCT 2570 3942 TCGCCCGGCGGAGCGGCCTC 2571 3943 CGCCCGGCGGAGCGGCCTCC 2572 3944 GCCCGGCGGAGCGGCCTCCC 2573 3945 CCCGGCGGAGCGGCCTCCCC 2574 3946 CCGGCGGAGCGGCCTCCCCA 2575 3947 CGGCGGAGCGGCCTCCCCAA 2576 3948 GGCGGAGCGGCCTCCCCAAG 2577 3949 GCGGAGCGGCCTCCCCAAGG 2578 3950 CGGAGCGGCCTCCCCAAGGT 2579 3951 GGAGCGGCCTCCCCAAGGTC 2580 3952 GAGCGGCCTCCCCAAGGTCA 2581 3953 AGCGGCCTCCCCAAGGTCAT 2582 3954 GCGGCCTCCCCAAGGTCATG 2583 3955 CGGCCTCCCCAAGGTCATGA 2584 3956 CTCTCGGGTCCCGGCCTCGC 2555 3957 TCTCTCGGGTCCCGGCCTCG 2554 3958 TTCTCTCGGGTCCCGGCCTC 2553 3959 CTTCTCTCGGGTCCCGGCCT 2552 3960 CCTTCTCTCGGGTCCCGGCC 2551 3961 CCCTTCTCTCGGGTCCCGGC 2550 3962 TCCCTTCTCTCGGGTCCCGG 2549 3963 GTCCCTTCTCTCGGGTCCCG 2548 3964 CGTCCCTTCTCTCGGGTCCC 2547 3965 GCGTCCCTTCTCTCGGGTCC 2546 3966 GGCGTCCCTTCTCTCGGGTC 2545 3967 CGGCGTCCCTTCTCTCGGGT 2544 3968 GCGGCGTCCCTTCTCTCGGG 2543 3969 GGCGGCGTCCCTTCTCTCGG 2542 3970 CGGCGGCGTCCCTTCTCTCG 2541 3971 CCGGCGGCGTCCCTTCTCTC 2540 3972 CCCGGCGGCGTCCCTTCTCT 2539 3973 GCCCGGCGGCGTCCCTTCTC 2538 3974 GGCCCGGCGGCGTCCCTTCT 2537 3975 AGGCCCGGCGGCGTCCCTTC 2536 3976 CAGGCCCGGCGGCGTCCCTT 2535 3977 CCAGGCCCGGCGGCGTCCCT 2534 3978 TCCAGGCCCGGCGGCGTCCC 2533 3979 CTCCAGGCCCGGCGGCGTCC 2532 3980 GCTCCAGGCCCGGCGGCGTC 2531 3981 AGCTCCAGGCCCGGCGGCGT 2530 3982 CAGCTCCAGGCCCGGCGGCG 2529 3983 CCAGCTCCAGGCCCGGCGGC 2528 3984 CCCAGCTCCAGGCCCGGCGG 2527 3985 CCCCAGCTCCAGGCCCGGCG 2526 3986 GCCCCAGCTCCAGGCCCGGC 2525 3987 AGCCCCAGCTCCAGGCCCGG 2524 3988 GAGCCCCAGCTCCAGGCCCG 2523 3989 CGAGCCCCAGCTCCAGGCCC 2522 3990 CCGAGCCCCAGCTCCAGGCC 2521 3991 CCCGAGCCCCAGCTCCAGGC 2520 3992 CCCCGAGCCCCAGCTCCAGG 2519 3993 ACCCCGAGCCCCAGCTCCAG 2518 3994 CACCCCGAGCCCCAGCTCCA 2517 3995 GCACCCCGAGCCCCAGCTCC 2516 3996 GGCACCCCGAGCCCCAGCTC 2515 3997 GGGCACCCCGAGCCCCAGCT 2514 3998 GGGGCACCCCGAGCCCCAGC 2513 3999 AGGGGCACCCCGAGCCCCAG 2512 4000 CAGGGGCACCCCGAGCCCCA 2511 4001 GCAGGGGCACCCCGAGCCCC 2510 4002 CGCAGGGGCACCCCGAGCCC 2509 4003 CCGCAGGGGCACCCCGAGCC 2508 4004 CCCGCAGGGGCACCCCGAGC 2507 4005 GCCCGCAGGGGCACCCCGAG 2506 4006 GGCCCGCAGGGGCACCCCGA 2505 4007 CGGCCCGCAGGGGCACCCCG 2504 4008 ACGGCCCGCAGGGGCACCCC 2503 4009 CACGGCCCGCAGGGGCACCC 2502 4010 CCACGGCCCGCAGGGGCACC 2501 4011 CCCACGGCCCGCAGGGGCAC 2500 4012 CCCCACGGCCCGCAGGGGCA 2499 4013 GCCCCACGGCCCGCAGGGGC 2498 4014 AGCCCCACGGCCCGCAGGGG 2497 4015 GAGCCCCACGGCCCGCAGGG 2496 4016 GGAGCCCCACGGCCCGCAGG 2495 4017 GGGAGCCCCACGGCCCGCAG 2494 4018 GGGGAGCCCCACGGCCCGCA 2493 4019 CGGGGAGCCCCACGGCCCGC 2492 4020 CCGGGGAGCCCCACGGCCCG 2491 4021 CCCGGGGAGCCCCACGGCCC 2490 4022 GCCCGGGGAGCCCCACGGCC 2489 4023 CGCCCGGGGAGCCCCACGGC 2488 4024 GCGCCCGGGGAGCCCCACGG 2487 4025 AGCGCCCGGGGAGCCCCACG 2486 4026 CAGCGCCCGGGGAGCCCCAC 2485 4027 CCAGCGCCCGGGGAGCCCCA 2484 4028 CCCAGCGCCCGGGGAGCCCC 2483 4029 GCCCAGCGCCCGGGGAGCCC 2482 4030 GGCCCAGCGCCCGGGGAGCC 2481 4031 TGGCCCAGCGCCCGGGGAGC 2480 4032 CTGGCCCAGCGCCCGGGGAG 2479 4033 GCTGGCCCAGCGCCCGGGGA 2478 4034 CGCTGGCCCAGCGCCCGGGG 2477 4035 TCGCTGGCCCAGCGCCCGGG 2476 4036 CTCGCTGGCCCAGCGCCCGG 2475 4037 CCTCGCTGGCCCAGCGCCCG 2474 4038 GCCTCGCTGGCCCAGCGCCC 2473 4039 TGCCTCGCTGGCCCAGCGCC 2472 4040 CTGCCTCGCTGGCCCAGCGC 2471 4041 GCTGCCTCGCTGGCCCAGCG 2470 4042 GGCTGCCTCGCTGGCCCAGC 2469 4043 CGGCTGCCTCGCTGGCCCAG 2468 4044 CCGGCTGCCTCGCTGGCCCA 2467 4045 GCCGGCTGCCTCGCTGGCCC 2466 4046 CGCCGGCTGCCTCGCTGGCC 2465 4047 CCGCCGGCTGCCTCGCTGGC 2464 4048 TCCGCCGGCTGCCTCGCTGG 2463 4049 CTCCGCCGGCTGCCTCGCTG 2462 4050 CCTCCGCCGGCTGCCTCGCT 2461 4051 ACCTCCGCCGGCTGCCTCGC 2460 4052 CACCTCCGCCGGCTGCCTCG 2459 4053 GCACCTCCGCCGGCTGCCTC 2458 4054 GGCACCTCCGCCGGCTGCCT 2457 4055 GGGCACCTCCGCCGGCTGCC 2456 4056 GGGGCACCTCCGCCGGCTGC 2455 4057 CGGGGCACCTCCGCCGGCTG 2454 4058 CCGGGGCACCTCCGCCGGCT 2453 4059 CCCGGGGCACCTCCGCCGGC 2452 4060 CCCCGGGGCACCTCCGCCGG 2451 4061 ACCCCGGGGCACCTCCGCCG 2450 4062 AACCCCGGGGCACCTCCGCC 2449 4063 CAACCCCGGGGCACCTCCGC 2448 4064 CCAACCCCGGGGCACCTCCG 2447 4065 TCCAACCCCGGGGCACCTCC 2446 4066 CTCCAACCCCGGGGCACCTC 2445 4067 TCTCCAACCCCGGGGCACCT 2444 4068 TTCTCCAACCCCGGGGCACC 2443 4069 TTTCTCCAACCCCGGGGCAC 2442 4070 CTTTCTCCAACCCCGGGGCA 2441 4071 TCTTTCTCCAACCCCGGGGC 2440 4072 GTCTTTCTCCAACCCCGGGG 2439 4073 AGTCTTTCTCCAACCCCGGG 2438 4074 GAGTCTTTCTCCAACCCCGG 2437 4075 CGAGTCTTTCTCCAACCCCG 2436 4076 GCGAGTCTTTCTCCAACCCC 2435 4077 GGCGAGTCTTTCTCCAACCC 2434 4078 CGGCGAGTCTTTCTCCAACC 2433 4079 GCGGCGAGTCTTTCTCCAAC 2432 4080 CGCGGCGAGTCTTTCTCCAA 2431 4081 CCGCGGCGAGTCTTTCTCCA 2430 4082 GCCGCGGCGAGTCTTTCTCC 2429 4083 GGCCGCGGCGAGTCTTTCTC 2428 4084 CATTCTGGCGGCGCCGCGGCTCGCG 2730 4085 ATTCTGGCGGCGCCGCGGCT 2731 4086 TTCTGGCGGCGCCGCGGCTC 2732 4087 TCTGGCGGCGCCGCGGCTCG 2733 4088 CTGGCGGCGCCGCGGCTCGC 2734 4089 TGGCGGCGCCGCGGCTCGCG 2735 4090 GGCGGCGCCGCGGCTCGCGC 2736 4091 GCGGCGCCGCGGCTCGCGCC 2737 4092 CGGCGCCGCGGCTCGCGCCC 2738 4093 GGCGCCGCGGCTCGCGCCCC 2739 4094 GCGCCGCGGCTCGCGCCCCG 2740 4095 CGCCGCGGCTCGCGCCCCGG 2741 4096 GCCGCGGCTCGCGCCCCGGC 2742 4097 CCGCGGCTCGCGCCCCGGCC 2743 4098 CGCGGCTCGCGCCCCGGCCC 2744 4099 GCGGCTCGCGCCCCGGCCCG 2745 4100 CGGCTCGCGCCCCGGCCCGA 2746 4101 GGCTCGCGCCCCGGCCCGAC 2747 4102 GCTCGCGCCCCGGCCCGACC 2748 4103 CCATTCTGGCGGCGCCGCGG 2729 4104 TCCATTCTGGCGGCGCCGCG 2728 4105 CTCCATTCTGGCGGCGCCGC 2727 4106 CCTCCATTCTGGCGGCGCCG 2726 4107 TCCTCCATTCTGGCGGCGCC 2725 4108 CTCCTCCATTCTGGCGGCGC 2724 4109 GCTCCTCCATTCTGGCGGCG 2723 4110 CGCTCCTCCATTCTGGCGGC 2722 4111 CCGCTCCTCCATTCTGGCGG 2721 4112 CCCGCTCCTCCATTCTGGCG 2720 4113 TCCCGCTCCTCCATTCTGGC 2719 4114 CTCCCGCTCCTCCATTCTGG 2718 4115 GCTCCCGCTCCTCCATTCTG 2717 4116 TGCTCCCGCTCCTCCATTCT 2716 4117 CTGCTCCCGCTCCTCCATTC 2715 4118 CCTGCTCCCGCTCCTCCATT 2714 4119 TCCTGCTCCCGCTCCTCCAT 2713 4120 TTCCTGCTCCCGCTCCTCCA 2712 4121 CTTCCTGCTCCCGCTCCTCC 2711 4122 ACTTCCTGCTCCCGCTCCTC 2710 4123 CACTTCCTGCTCCCGCTCCT 2709 4124 CCACTTCCTGCTCCCGCTCC 2708 4125 GCCACTTCCTGCTCCCGCTC 2707 4126 GGCCACTTCCTGCTCCCGCT 2706 4127 CGGCCACTTCCTGCTCCCGC 2705 4128 TCGGCCACTTCCTGCTCCCG 2704 4129 CTCGGCCACTTCCTGCTCCC 2703 4130 GCTCGGCCACTTCCTGCTCC 2702 4131 CGCTCGGCCACTTCCTGCTC 2701 4132 CCGCTCGGCCACTTCCTGCT 2700 4133 CCCGCTCGGCCACTTCCTGC 2699 4134 GCCCGCTCGGCCACTTCCTG 2698 4135 GGCCCGCTCGGCCACTTCCT 2697 4136 AGGCCCGCTCGGCCACTTCC 2696 4137 CAGGCCCGCTCGGCCACTTC 2695 4138 CCAGGCCCGCTCGGCCACTT 2694 4139 CCCAGGCCCGCTCGGCCACT 2693 4140 GCCCAGGCCCGCTCGGCCAC 2692 4141 CGCCCAGGCCCGCTCGGCCA 2691 4142 CCGCCCAGGCCCGCTCGGCC 2690 4143 CCCGCCCAGGCCCGCTCGGC 2689 4144 CCCCGCCCAGGCCCGCTCGG 2688 4145 TCCCCGCCCAGGCCCGCTCG 2687 4146 CTCCCCGCCCAGGCCCGCTC 2686 4147 CCTCCCCGCCCAGGCCCGCT 2685 4148 CCCTCCCCGCCCAGGCCCGC 2684 4149 GCCCTCCCCGCCCAGGCCCG 2683 4150 CGCCCTCCCCGCCCAGGCCC 2682 4151 GCGCCCTCCCCGCCCAGGCC 2681 4152 CGCGCCCTCCCCGCCCAGGC 2680 4153 CCGCGCCCTCCCCGCCCAGG 2679 4154 CCCGCGCCCTCCCCGCCCAG 2678 4155 CCCCGCGCCCTCCCCGCCCA 2677 4156 GCCCCGCGCCCTCCCCGCCC 2676 4157 CGCCCCGCGCCCTCCCCGCC 2675 4158 GCGCCCCGCGCCCTCCCCGC 2674 4159 CGCGCCCCGCGCCCTCCCCG 2673 4160 GCGCGCCCCGCGCCCTCCCC 2672 4161 CGCGCGCCCCGCGCCCTCCC 2671 4162 CCGCGCGCCCCGCGCCCTCC 2670 4163 CCCGCGCGCCCCGCGCCCTC 2669 4164 GCCCGCGCGCCCCGCGCCCT 2668 4165 GGCCCGCGCGCCCCGCGCCC 2667 4166 GGGCCCGCGCGCCCCGCGCC 2666 4167 CGGGCCCGCGCGCCCCGCGC 2665 4168 CCGGGCCCGCGCGCCCCGCG 2664 4169 GCCGGGCCCGCGCGCCCCGC 2663 4170 GGCCGGGCCCGCGCGCCCCG 2662 4171 TGGCCGGGCCCGCGCGCCCC 2661 4172 TTGGCCGGGCCCGCGCGCCC 2660 4173 CTTGGCCGGGCCCGCGCGCC 2659 4174 CCTTGGCCGGGCCCGCGCGC 2658 4175 CCCTTGGCCGGGCCCGCGCG 2657 4176 TCCCTTGGCCGGGCCCGCGC 2656 4177 CTCCCTTGGCCGGGCCCGCG 2655 4178 CCTCCCTTGGCCGGGCCCGC 2654 4179 CCCTCCCTTGGCCGGGCCCG 2653 4180 GCCCTCCCTTGGCCGGGCCC 2652 4181 CGCCCTCCCTTGGCCGGGCC 2651 4182 CCGCCCTCCCTTGGCCGGGC 2650 4183 GCCGCCCTCCCTTGGCCGGG 2649 4184 GGCCGCCCTCCCTTGGCCGG 2648 4185 GGGCCGCCCTCCCTTGGCCG 2647 4186 GGGGCCGCCCTCCCTTGGCC 2646 4187 TGGGGCCGCCCTCCCTTGGC 2645 4188 GTGGGGCCGCCCTCCCTTGG 2644 4189 CGTGGGGCCGCCCTCCCTTG 2643 4190 GCGTGGGGCCGCCCTCCCTT 2642 4191 GGCGTGGGGCCGCCCTCCCT 2641 4192 CGGCGTGGGGCCGCCCTCCC 2640 4193 CCGGCGTGGGGCCGCCCTCC 2639 4194 CCCGGCGTGGGGCCGCCCTC 2638 4195 GCCCGGCGTGGGGCCGCCCT 2637 4196 CGCCCGGCGTGGGGCCGCCC 2636 4197 GCGCCCGGCGTGGGGCCGCC 2635 4198 GGCGCCCGGCGTGGGGCCGC 2634 4199 CGGCGCCCGGCGTGGGGCCG 2633 4200 CCGGCGCCCGGCGTGGGGCC 2632 4201 CCCGGCGCCCGGCGTGGGGC 2631 4202 CCCCGGCGCCCGGCGTGGGG 2630 4203 CCCCCGGCGCCCGGCGTGGG 2629 4204 ACCCCCGGCGCCCGGCGTGG 2628 4205 CACCCCCGGCGCCCGGCGTG 2627 4206 GCACCCCCGGCGCCCGGCGT 2626 4207 TGCACCCCCGGCGCCCGGCG 2625 4208 CTGCACCCCCGGCGCCCGGC 2624 4209 CCTGCACCCCCGGCGCCCGG 2623 4210 GCCTGCACCCCCGGCGCCCG 2622 4211 AGCCTGCACCCCCGGCGCCC 2621 4212 CAGCCTGCACCCCCGGCGCC 2620 4213 GCAGCCTGCACCCCCGGCGC 2619 4214 GGCAGCCTGCACCCCCGGCG 2618 4215 CGGCAGCCTGCACCCCCGGC 2617 4216 CCGGCAGCCTGCACCCCCGG 2616 4217 GCCGGCAGCCTGCACCCCCG 2615 4218 GGCCGGCAGCCTGCACCCCC 2614 4219 GGGCCGGCAGCCTGCACCCC 2613 4220 GGGGCCGGCAGCCTGCACCC 2612 4221 TGGGGCCGGCAGCCTGCACC 2611 4222 CTGGGGCCGGCAGCCTGCAC 2610 4223 GCTGGGGCCGGCAGCCTGCA 2609 4224 GGCTGGGGCCGGCAGCCTGC 2608 4225 AGGCTGGGGCCGGCAGCCTG 2607 4226 GAGGCTGGGGCCGGCAGCCT 2606 4227 GGAGGCTGGGGCCGGCAGCC 2605 4228 CACCGGGCCGCCGCGTCCGGGCGCG 2838 4229 ACCGGGCCGCCGCGTCCGGG 2839 4230 CCGGGCCGCCGCGTCCGGGC 2840 4231 CGGGCCGCCGCGTCCGGGCG 2841 4232 GGGCCGCCGCGTCCGGGCGC 2842 4233 GGCCGCCGCGTCCGGGCGCG 2843 4234 GCCGCCGCGTCCGGGCGCGA 2844 4235 CCGCCGCGTCCGGGCGCGAG 2845 4236 CGCCGCGTCCGGGCGCGAGC 2846 4237 GCCGCGTCCGGGCGCGAGCG 2847 4238 CCGCGTCCGGGCGCGAGCGC 2848 4239 CGCGTCCGGGCGCGAGCGCG 2849 4240 GCGTCCGGGCGCGAGCGCGG 2850 4241 CGTCCGGGCGCGAGCGCGGG 2851 4242 GTCCGGGCGCGAGCGCGGGC 2852 4243 TCCGGGCGCGAGCGCGGGCC 2853 4244 CCGGGCGCGAGCGCGGGCCT 2854 4245 CGGGCGCGAGCGCGGGCCTA 2855 4246 GGGCGCGAGCGCGGGCCTAG 2856 4247 GGCGCGAGCGCGGGCCTAGC 2857 4248 GCGCGAGCGCGGGCCTAGCC 2858 4249 CGCGAGCGCGGGCCTAGCCG 2859 4250 GCGAGCGCGGGCCTAGCCGG 2860 4251 CGAGCGCGGGCCTAGCCGGG 2861 4252 GAGCGCGGGCCTAGCCGGGC 2862 4253 AGCGCGGGCCTAGCCGGGCC 2863 4254 GCGCGGGCCTAGCCGGGCCG 2864 4255 CGCGGGCCTAGCCGGGCCGC 2865 4256 GCGGGCCTAGCCGGGCCGCG 2866 4257 CGGGCCTAGCCGGGCCGCGG 2867 4258 GGGCCTAGCCGGGCCGCGGC 2868 4259 GGCCTAGCCGGGCCGCGGCC 2869 4260 GCCTAGCCGGGCCGCGGCCT 2870 4261 CCTAGCCGGGCCGCGGCCTC 2871 4262 CTAGCCGGGCCGCGGCCTCC 2872 4263 TAGCCGGGCCGCGGCCTCCG 2873 4264 AGCCGGGCCGCGGCCTCCGG 2874 4265 GCCGGGCCGCGGCCTCCGGC 2875 4266 CCGGGCCGCGGCCTCCGGCG 2876 4267 CGGGCCGCGGCCTCCGGCGC 2877 4268 GGGCCGCGGCCTCCGGCGCC 2878 4269 GGCCGCGGCCTCCGGCGCCC 2879 4270 GCCGCGGCCTCCGGCGCCCG 2880 4271 CCGCGGCCTCCGGCGCCCGC 2881 4272 CGCGGCCTCCGGCGCCCGCC 2882 4273 GCGGCCTCCGGCGCCCGCCG 2883 4274 CGGCCTCCGGCGCCCGCCGC 2884 4275 GGCCTCCGGCGCCCGCCGCT 2885 4276 GCCTCCGGCGCCCGCCGCTC 2886 4277 CCTCCGGCGCCCGCCGCTCC 2887 4278 CTCCGGCGCCCGCCGCTCCG 2888 4279 TCCGGCGCCCGCCGCTCCGC 2889 4280 CCGGCGCCCGCCGCTCCGCA 2890 4281 CGGCGCCCGCCGCTCCGCAT 2891 4282 GGCGCCCGCCGCTCCGCATC 2892 4283 GCGCCCGCCGCTCCGCATCC 2893 4284 CGCCCGCCGCTCCGCATCCC 2894 4285 GCCCGCCGCTCCGCATCCCC 2895 4286 CCCGCCGCTCCGCATCCCCG 2896 4287 CCGCCGCTCCGCATCCCCGC 2897 4288 CGCCGCTCCGCATCCCCGCG 2898 4289 GCCGCTCCGCATCCCCGCGG 2899 4290 CCGCTCCGCATCCCCGCGGG 2900 4291 CGCTCCGCATCCCCGCGGGC 2901 4292 GCTCCGCATCCCCGCGGGCC 2902 4293 CTCCGCATCCCCGCGGGCCG 2903 4294 TCCGCATCCCCGCGGGCCGG 2904 4295 CCGCATCCCCGCGGGCCGGC 2905 4296 CGCATCCCCGCGGGCCGGCG 2906 4297 GCATCCCCGCGGGCCGGCGC 2907 4298 CATCCCCGCGGGCCGGCGCT 2908 4299 ATCCCCGCGGGCCGGCGCTG 2909 4300 TCCCCGCGGGCCGGCGCTGG 2910 4301 CCCCGCGGGCCGGCGCTGGG 2911 4302 CCCGCGGGCCGGCGCTGGGC 2912 4303 CCGCGGGCCGGCGCTGGGCG 2913 4304 CGCGGGCCGGCGCTGGGCGG 2914 4305 GCGGGCCGGCGCTGGGCGGG 2915 4306 CGGGCCGGCGCTGGGCGGGG 2916 4307 GGGCCGGCGCTGGGCGGGGC 2917 4308 GGCCGGCGCTGGGCGGGGCC 2918 4309 GCCGGCGCTGGGCGGGGCCG 2919 4310 CCGGCGCTGGGCGGGGCCGG 2920 4311 CGGCGCTGGGCGGGGCCGGG 2921 4312 GGCGCTGGGCGGGGCCGGGC 2922 4313 GCGCTGGGCGGGGCCGGGCT 2923 4314 CGCTGGGCGGGGCCGGGCTG 2924 4315 GCTGGGCGGGGCCGGGCTGG 2925 4316 CTGGGCGGGGCCGGGCTGGA 2926 4317 TCACCGGGCCGCCGCGTCCG 2837 4318 CTCACCGGGCCGCCGCGTCC 2836 4319 ACTCACCGGGCCGCCGCGTC 2835 4320 GACTCACCGGGCCGCCGCGT 2834 4321 GGACTCACCGGGCCGCCGCG 2833 4322 GGGACTCACCGGGCCGCCGC 2832 4323 GGGGACTCACCGGGCCGCCG 2831 4324 CGGGGACTCACCGGGCCGCC 2830 4325 GCGGGGACTCACCGGGCCGC 2829 4326 GGCGGGGACTCACCGGGCCG 2828 4327 GGGCGGGGACTCACCGGGCC 2827 4328 CGGGCGGGGACTCACCGGGC 2826 4329 GCGGGCGGGGACTCACCGGG 2825 4330 GGCGGGCGGGGACTCACCGG 2824 4331 CGGCGGGCGGGGACTCACCG 2823 4332 ACGGCGGGCGGGGACTCACC 2822 4333 CACGGCGGGCGGGGACTCAC 2821 4334 CCACGGCGGGCGGGGACTCA 2820 4335 GCCACGGCGGGCGGGGACTC 2819 4336 GGCCACGGCGGGCGGGGACT 2818 4337 CGGCCACGGCGGGCGGGGAC 2817 4338 CACATCCGCCTCCGCCGCCCGG 3160

Hot Zones (Relative upstream location to gene start site)  1-350  700-1100 1500-1650 1750-3650

Examples

In FIG. 33, In MCF7 (human mammary breast cell line), AKT4 (169) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The AKT sequence AKT4 (169) fits the independent and dependent DNAi motif claims.

The secondary structure for AKT4 (169) is shown in FIG. 34.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11965) CGGCAGGACCGAGCGCGGCAGGCGGCTGGCCCAGCGCACGCAGCGCGGCC CGAAGACGGGAGCAGGCGGCCGAGCACCGAGCGCTGGGCACCGGGCACCG AGCGGCGGCGGCACGCGAGGCCCGGCCCCGAGCAGCGCCCCCGCCCGCCG CGGCCTCCAGCCCGGCCCCGCCCAGCGCCGGCCCGCGGGGATGCGGAGCG GCGGGCGCCGGAGGCCGCGGCCCGGCTAGGCCCGCGCTCGCGCCCGGACG CGGCGGCCCGGTGAGTCCCCGCCCGCCGTGGCCGCCCGGGCCTGGATTTC CTCCCCGCGGGCCGGGCCGCTTTGTTCGCGGCCGGTCGGGCCGGGGCGCG AGCCGCGGCGCCGCCAGAATGGAGGAGCGGGAGCAGGAAGTGGCCGAGCG GGCCTGGGCGGGGAGGGCGCGGGGCGCGCGGGCCCGGCCAAGGGAGGGCG GCCCCACGCCGGGCGCCGGGGGTGCAGGCTGCCGGCCCCAGCCTCCCTCA TGACCTTGGGGAGGCCGCTCCGCCGGGCGAGGCCGGGACCCGAGAGAAGG GACGCCGCCGGGCCTGGAGCTGGGGCTCGGGGTGCCCCTGCGGGCCGTGG GGCTCCCCGGGCGCTGGGCCAGCGAGGCAGCCGGCGGAGGTGCCCCGGGG TTGGAGAAAGACTCGCCGCGGCCGGCCTTCAAGTTTGTGGGAGGGCCCCG GAAGGAGACTTCGTTTCCCACGGACGAAAAGTTGTACGTGGTGGCGGGGT ACCCAGGCTAGCCACAAAGGACTGTGACCCTCCTGGGCCCCGGAACTGCT TCCTGTCTTGGGTGGGCCCTGGAGGTCCTGCCCGCCCATCCCAGAGGCCA AGGCTTGGAGGGCAGCTGGGGCTTGCCCCTTAGATTGAGTATCCTGGGGC GCTAGCGAGCTTGGTCCTGTCGGGACGGCCTCTGAGTGCTGCCTTGGTCA GCGGGTGAGCTTGGGCCCCTGCTCTGCAGCCAGAGGCCGCCCCACATTCA CTCCTGGGTCTCTCGGCCTTGCTCCAGGTGGCCACTTCTTGACTGCTTTG AGTCCCTCATCCGAGCGAAGGGCGGACGGAGTCCGTTGGTGGGGGTCCGG TTGCCTCTCCCGGGAGCTGTGTAGACTTCTCATACACCAGGGTTCTGGAG GCAGATGGAGGAGCCCTTTCGAAAACAGAGTATTTTTTTTTAAGTTGTGA CTTAATAATAGTAGCAAGAATATGTGCTTATGGTAAAGGCAGGCGGCAGG TACGGAGGCTGTGGGAAGTCGGGGTCCCTCCGCCCCCACAGGCAGCCCTG TGCTGGCCTGGTGTATACGTTTCTGTGCAGACGTACACCACCCTGTGTGA GCACAGATGTATTTTTACACATGGCTCTGGACAGCTGTCTGACTCTGTCA GCAGCAGGCCTTGGAGGGGCTCAGGCCCGTGTGGGGGTGGGGGGACATCC AGAGGTCTTTGAGTCCAGCCCTCTGCCTCCAGGCCACGCCCACTCAGTGT CGTCAGAGCCCCCTGTGCCTGAGGCGTGCGCGGCTCGGAGCCCTGCCCTC GGAGTCCTGCGGTGCCTTCCTCGAGTCTGGCCTGCTTTCCATCCTGCTAA GTACTTGGGGCATTTCCCTCTTTGGGTAAGGTGTGGTCTTCCCTGTCCTG GCATTAGACACAAGGCAGTGGGCCTTCCTGCCATTCTAAGTGTAGCTTAA GACAATCAGTGCAAAGCAACCCTTTGTGGGTGTCCAGCCCTTGCCTCGGG AGGCCAGAAAGGTGGCCTGGGGGGAGAGCGTCTAAGCTGGCTGTGGAAAG ACCCATGTTGGGATCCATTCCACAGAGGTCGTCAGGGGTCTCTGCCTGGC CTGGAGGTCCCAGAGAGGACCCTCCTCCCCTCAGGAAGGCCCATCTGGAA GGGTAGCAGAGGACTGCTCACAGGAAGAGCATGCGAAGTGCTCTTTCTGG GGATGCCTGTAGTTGGTGATGTGGGAACTGGGTTTTGAGGGATGCCTAGG AGTTCATCCATCAGAGGGGAAATGAGGAAGCCATGCAGGATCAATGGATA AAGTGTGCTCAGGTGAGGGTTGGCTGGTGGGCCGCTGCAGGGCGGGGGCC TGTCCAGTGCTCCCCCACTTACTTGCTGCCTCCCGACTGCTGTAATTATG GGTCTGTAACCACCCTGGACTGGGTGCTCCTCACTGACGGACTTGTCTGA ACCTCTCTTTGTCTCCAGCGCCCAGCACTGGGCCTGGCAAAACCTGAGAC GCCCGGTACATGTTGGCCAAATGAATGAACCAGATTCAGACCGGCAGGGG CGCTGTGGTTTAGGAGGGGCCTGGGGTTTCTCCCAGGAGGTTTTTGGGCT TGCGCTGGAGGGCTCTGGACTCCCGTTTGCGCCAGTGGCCTGCATCCTGG TCCTGTCTTCCTCATGTTTGAATTTCTTTGCTTTCCTAGTCTGGGGAGCA GGGAGGAGCCCTGTGCCCTGTCCCAGGATCCATGGGTAGGAACACCATGG ACAGGGAGAGCAAACGGGGCCATCTGTCACCAGGGGCTTAGGGAAGGCCG AGCCAGCCTGGGTCAAAGAAGTCAAAGGGGCTGCCTGGAGGAGGCAGCCT GTCAGCTGGTGCATCAGGTTAGGGAGGCTGGGAAGGCCTTTTGGGGATGG GGGTGATTTGTCCAACGGCTGGGGGAGGTGGGAATGGGGAGGTGAGCAAG GCAGCAGCTCTCAGGGCCTGGCTGTTGCGGGTGGTGGTGGCAGGGGCTGG AGGCTCTAAGCCTAGAATAAGGAGAGGCCCAGGTCCAGGGAACTGTGTTC AATTACATGGATTTGACACTTGGCAGCCCTGAGTGTTTTGGGGAGAGGGA AGGCAGGCGGGCAGATGGGGGTCAGAGAGCTTAGAGGGATGGCAGCCCAC CTGGGAAGGCAGGTGCGGGTGGAGCCCCCAGGCACGTGCAGTGGGTCTCT GGCTCACCCAGGGCGAGGAGCTGCCCTTAGCCAGGCGTGGCCTCACATTC AGCTTCCTTTGCTTCTCCCAGAGGCTGTGGCCAGGCCAGCTGGGCTCGGG GAGCGCCAGCCTGAGAGGAGCGCGTGAGCGTCGCGGGAGCCTCGGGCACC ATG

17) CRAF. RAF proto-oncogene serine/threonine-protein kinase also known as proto-oncogene c-RAF or simply c-Raf or even Raf-1 is an enzyme is encoded by the RAF1 gene. The c-Raf protein is part of the ERK1/2 pathway as a MAP kinase kinase kinase (MAP3K) that functions downstream of the Ras subfamily of membrane associated GTPases.

Elevated C-Raf mRNA or protein levels have been identified in AML, head and neck cancer, prostate cancer and ovarian cancer (Schmidt et al., Leuk Res. 1994; 18:409-13, Riva et al., Eur J Cancer B Oral Oncol. 1995; 31B:384-91, Muhkerjhee et al., Prostate. 2005; 64:101-7). In ovarian cancer cell lines, antisense oligodeoxynucleotides (ODNs) inhibited cell proliferation in vitro (McPhillips et al., Br J Cancer. 2001; 85:1753-8) with similar results seen in lines derived from lung, cervical, prostate and colon carcinomas showed the same phenomenon.

Inhibiting cRAF may be useful against diabetic retinopathy, one of the leading causes of blindness A c-RAF inhibitor (iCo-007) is being developed for the treatment of various eye diseases that occur as complications of diabetes. In patients with diffuse diabetic macular edema presented positive results from the Phase 1 study showing that subjects tolerated iCo-007 well. In this study, a number of individuals exhibited a decrease of central macular edema compared to baseline using an analytical method called optical coherence tomography prompting the initiation of a Phase 2 study on iCo-007 in patients with diabetic macular edema.

Hereditary gain-of-function mutations of c-Raf are implicated in some rare, but severe syndromes. Mutation of c-Raf is one of the possible causes of Noonan syndrome: affected individuals have congenital heart defects, short and dysmorhic stature and several other deformities. Similar mutations in c-Raf can also cause a related condition, termed LEOPARD syndrome (Lentigo, Electrocardiographic abnormalities, Ocular hypertelorism, Pulmonary stenosis, Abnormal genitalia, Retarded growth, Deafness), with a complex association of defects.

Protein: c-Raf Gene: RAF1 (Homo sapiens, chromosome 3, 12625100-12705700 [NCBI Reference Sequence: NC_(—)000003.11]; start site location: 12660220; strand: negative)

Gene Identification GeneID 5894 HGNC 9829 HPRD 01265 MIM 164760

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 4339 GCGCGAGCCCTACTGGCAGTCG 25996 4462 CGGGGCGTGGCCTAGCGATCTGGTGGCCG 26073 4517 TTTCGAAGCTGAAGAGGTTAGGCGACG 26106 4519 CGACGCTGACTTGCTTTCAGGAG 26127 4533 AATCGAGAAGAACCGGCTTTCGG 26161 4556 CTTTGACGCGTCCTCTCCGGGC 26295 4585 CGGCTCCGCCACTTGACAGCTATGTGG 26334 4605 AGGCGGAGATTGCGGTGAGCCGAAATCGCG 27188 4609 AGGCCGCCCCAACGTCCTGTCGTTCGGCGG 25618 4677 TCTCGCCCGCTCCTCCTCCCCGCGGCGGGTG 25653 4745 CGGGAGGCGGTCACATTCGGCGCG 25690 4782 CGGAGCCCCGAGCAGCCCCCGCATCG 25730 4871 CGCGCTCCGCGCCTCAGGGCACGCGCC 25763 4960 AGCCGTTCCCGCCTCACAATCG 25840 4984 CCGCCATCTAAGATGGCGGCC 25876 5047 CGGGCGGCCCAGACGAGCGAGCCCTCG 25920 5110 CGTCCTCCCGACCTGCGACGCCACCGGC 25957

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 4339 GCGCGAGCCCTACTGGCAGTCG 25996 4340 CGCGAGCCCTACTGGCAGTC 25997 4341 GCGAGCCCTACTGGCAGTCG 25998 4342 CGAGCCCTACTGGCAGTCGA 25999 4343 GAGCCCTACTGGCAGTCGAC 26000 4344 AGCCCTACTGGCAGTCGACT 26001 4345 GCCCTACTGGCAGTCGACTT 26002 4346 CCCTACTGGCAGTCGACTTC 26003 4347 CCTACTGGCAGTCGACTTCT 26004 4348 CTACTGGCAGTCGACTTCTA 26005 4349 TACTGGCAGTCGACTTCTAA 26006 4350 ACTGGCAGTCGACTTCTAAC 26007 4351 CTGGCAGTCGACTTCTAACT 26008 4352 TGGCAGTCGACTTCTAACTT 26009 4353 GGCAGTCGACTTCTAACTTG 26010 4354 GCAGTCGACTTCTAACTTGG 26011 4355 CAGTCGACTTCTAACTTGGC 26012 4356 AGTCGACTTCTAACTTGGCT 26013 4357 GTCGACTTCTAACTTGGCTC 26014 4358 TCGACTTCTAACTTGGCTCG 26015 4359 CGACTTCTAACTTGGCTCGG 26016 4360 GACTTCTAACTTGGCTCGGG 26017 4361 ACTTCTAACTTGGCTCGGGC 26018 4362 CTTCTAACTTGGCTCGGGCA 26019 4363 TTCTAACTTGGCTCGGGCAT 26020 4364 TCTAACTTGGCTCGGGCATC 26021 4365 CTAACTTGGCTCGGGCATCC 26022 4366 TAACTTGGCTCGGGCATCCA 26023 4367 AACTTGGCTCGGGCATCCAT 26024 4368 ACTTGGCTCGGGCATCCATC 26025 4369 CTTGGCTCGGGCATCCATCG 26026 4370 TTGGCTCGGGCATCCATCGC 26027 4371 TGGCTCGGGCATCCATCGCT 26028 4372 GGCTCGGGCATCCATCGCTC 26029 4373 GCTCGGGCATCCATCGCTCT 26030 4374 CTCGGGCATCCATCGCTCTG 26031 4375 TCGGGCATCCATCGCTCTGG 26032 4376 CGGGCATCCATCGCTCTGGC 26033 4377 GGGCATCCATCGCTCTGGCC 26034 4378 GGCATCCATCGCTCTGGCCT 26035 4379 GCATCCATCGCTCTGGCCTG 26036 4380 CATCCATCGCTCTGGCCTGA 26037 4381 ATCCATCGCTCTGGCCTGAA 26038 4382 TCCATCGCTCTGGCCTGAAC 26039 4383 CCATCGCTCTGGCCTGAACT 26040 4384 CATCGCTCTGGCCTGAACTC 26041 4385 ATCGCTCTGGCCTGAACTCA 26042 4386 TCGCTCTGGCCTGAACTCAG 26043 4387 CGCTCTGGCCTGAACTCAGG 26044 4388 TGCGCGAGCCCTACTGGCAG 25995 4389 CTGCGCGAGCCCTACTGGCA 25994 4390 TCTGCGCGAGCCCTACTGGC 25993 4391 TTCTGCGCGAGCCCTACTGG 25992 4392 ATTCTGCGCGAGCCCTACTG 25991 4393 GATTCTGCGCGAGCCCTACT 25990 4394 CGATTCTGCGCGAGCCCTAC 25989 4395 CCGATTCTGCGCGAGCCCTA 25988 4396 TCCGATTCTGCGCGAGCCCT 25987 4397 CTCCGATTCTGCGCGAGCCC 25986 4398 TCTCCGATTCTGCGCGAGCC 25985 4399 CTCTCCGATTCTGCGCGAGC 25984 4400 GCTCTCCGATTCTGCGCGAG 25983 4401 GGCTCTCCGATTCTGCGCGA 25982 4402 CGGCTCTCCGATTCTGCGCG 25981 4403 CCGGCTCTCCGATTCTGCGC 25980 4404 ACCGGCTCTCCGATTCTGCG 25979 4405 CACCGGCTCTCCGATTCTGC 25978 4406 CCACCGGCTCTCCGATTCTG 25977 4407 GCCACCGGCTCTCCGATTCT 25976 4408 CGCCACCGGCTCTCCGATTC 25975 4409 ACGCCACCGGCTCTCCGATT 25974 4410 GACGCCACCGGCTCTCCGAT 25973 4411 CGACGCCACCGGCTCTCCGA 25972 4412 GCGACGCCACCGGCTCTCCG 25971 4413 TGCGACGCCACCGGCTCTCC 25970 4414 CTGCGACGCCACCGGCTCTC 25969 4415 CCTGCGACGCCACCGGCTCT 25968 4416 ACCTGCGACGCCACCGGCTC 25967 4417 GACCTGCGACGCCACCGGCT 25966 4418 CGACCTGCGACGCCACCGGC 25965 4419 CCGACCTGCGACGCCACCGG 25964 4420 CCCGACCTGCGACGCCACCG 25963 4421 TCCCGACCTGCGACGCCACC 25962 4422 CTCCCGACCTGCGACGCCAC 25961 4423 CCTCCCGACCTGCGACGCCA 25960 4424 TCCTCCCGACCTGCGACGCC 25959 4425 GTCCTCCCGACCTGCGACGC 25958 4426 CGTCCTCCCGACCTGCGACG 25957 4427 TCGTCCTCCCGACCTGCGAC 25956 4428 CTCGTCCTCCCGACCTGCGA 25955 4429 GCTCGTCCTCCCGACCTGCG 25954 4430 TGCTCGTCCTCCCGACCTGC 25953 4431 GTGCTCGTCCTCCCGACCTG 25952 4432 GGTGCTCGTCCTCCCGACCT 25951 4433 CGGTGCTCGTCCTCCCGACC 25950 4434 TCGGTGCTCGTCCTCCCGAC 25949 4435 CTCGGTGCTCGTCCTCCCGA 25948 4436 ACTCGGTGCTCGTCCTCCCG 25947 4437 GACTCGGTGCTCGTCCTCCC 25946 4438 CGACTCGGTGCTCGTCCTCC 25945 4439 TCGACTCGGTGCTCGTCCTC 25944 4440 CTCGACTCGGTGCTCGTCCT 25943 4441 CCTCGACTCGGTGCTCGTCC 25942 4442 CCCTCGACTCGGTGCTCGTC 25941 4443 GCCCTCGACTCGGTGCTCGT 25940 4444 AGCCCTCGACTCGGTGCTCG 25939 4445 GAGCCCTCGACTCGGTGCTC 25938 4446 CGAGCCCTCGACTCGGTGCT 25937 4447 GCGAGCCCTCGACTCGGTGC 25936 4448 AGCGAGCCCTCGACTCGGTG 25935 4449 GAGCGAGCCCTCGACTCGGT 25934 4450 CGAGCGAGCCCTCGACTCGG 25933 4451 ACGAGCGAGCCCTCGACTCG 25932 4452 GACGAGCGAGCCCTCGACTC 25931 4453 AGACGAGCGAGCCCTCGACT 25930 4454 CAGACGAGCGAGCCCTCGAC 25929 4455 CCAGACGAGCGAGCCCTCGA 25928 4456 CCCAGACGAGCGAGCCCTCG 25927 4457 GCCCAGACGAGCGAGCCCTC 25926 4458 GGCCCAGACGAGCGAGCCCT 25925 4459 CGGCCCAGACGAGCGAGCCC 25924 4460 GCGGCCCAGACGAGCGAGCC 25923 4461 GGCGGCCCAGACGAGCGAGC 25922 4462 CGGGGCGTGGCCTAGCGATCTGGTGGCCG 26073 4463 GGGGCGTGGCCTAGCGATCT 26074 4464 GGGCGTGGCCTAGCGATCTG 26075 4465 GGCGTGGCCTAGCGATCTGG 26076 4466 GCGTGGCCTAGCGATCTGGT 26077 4467 CGTGGCCTAGCGATCTGGTG 26078 4468 GTGGCCTAGCGATCTGGTGG 26079 4469 TGGCCTAGCGATCTGGTGGC 26080 4470 GGCCTAGCGATCTGGTGGCC 26081 4471 GCCTAGCGATCTGGTGGCCG 26082 4472 CCTAGCGATCTGGTGGCCGC 26083 4473 CTAGCGATCTGGTGGCCGCC 26084 4474 TAGCGATCTGGTGGCCGCCA 26085 4475 AGCGATCTGGTGGCCGCCAT 26086 4476 GCGATCTGGTGGCCGCCATT 26087 4477 CGATCTGGTGGCCGCCATTT 26088 4478 GATCTGGTGGCCGCCATTTC 26089 4479 ATCTGGTGGCCGCCATTTCG 26090 4480 TCTGGTGGCCGCCATTTCGA 26091 4481 CTGGTGGCCGCCATTTCGAA 26092 4482 TGGTGGCCGCCATTTCGAAG 26093 4483 GGTGGCCGCCATTTCGAAGC 26094 4484 GTGGCCGCCATTTCGAAGCT 26095 4485 TGGCCGCCATTTCGAAGCTG 26096 4486 GGCCGCCATTTCGAAGCTGA 26097 4487 GCCGCCATTTCGAAGCTGAA 26098 4488 CCGCCATTTCGAAGCTGAAG 26099 4489 CGCCATTTCGAAGCTGAAGA 26100 4490 GCCATTTCGAAGCTGAAGAG 26101 4491 CCATTTCGAAGCTGAAGAGG 26102 4492 CATTTCGAAGCTGAAGAGGT 26103 4493 CCGGGGCGTGGCCTAGCGAT 26072 4494 CCCGGGGCGTGGCCTAGCGA 26071 4495 CCCCGGGGCGTGGCCTAGCG 26070 4496 CCCCCGGGGCGTGGCCTAGC 26069 4497 GCCCCCGGGGCGTGGCCTAG 26068 4498 CGCCCCCGGGGCGTGGCCTA 26067 4499 CCGCCCCCGGGGCGTGGCCT 26066 4500 CCCGCCCCCGGGGCGTGGCC 26065 4501 CCCCGCCCCCGGGGCGTGGC 26064 4502 GCCCCGCCCCCGGGGCGTGG 26063 4503 GGCCCCGCCCCCGGGGCGTG 26062 4504 AGGCCCCGCCCCCGGGGCGT 26061 4505 CAGGCCCCGCCCCCGGGGCG 26060 4506 TCAGGCCCCGCCCCCGGGGC 26059 4507 CTCAGGCCCCGCCCCCGGGG 26058 4508 ACTCAGGCCCCGCCCCCGGG 26057 4509 AACTCAGGCCCCGCCCCCGG 26056 4510 GAACTCAGGCCCCGCCCCCG 26055 4511 TGAACTCAGGCCCCGCCCCC 26054 4512 CTGAACTCAGGCCCCGCCCC 26053 4513 CCTGAACTCAGGCCCCGCCC 26052 4514 GCCTGAACTCAGGCCCCGCC 26051 4515 GGCCTGAACTCAGGCCCCGC 26050 4516 TGGCCTGAACTCAGGCCCCG 26049 4517 TTTCGAAGCTGAAGAGGTTAGGCGACG 26105 4518 TTCGAAGCTGAAGAGGTTAG 26106 4519 CGACGCTGACTTGCTTTCAGGAG 26127 4520 GACGCTGACTTGCTTTCAGG 26128 4521 ACGCTGACTTGCTTTCAGGA 26129 4522 CGCTGACTTGCTTTCAGGAG 26130 4523 GCGACGCTGACTTGCTTTCA 26126 4524 GGCGACGCTGACTTGCTTTC 26125 4525 AGGCGACGCTGACTTGCTTT 26124 4526 TAGGCGACGCTGACTTGCTT 26123 4527 TTAGGCGACGCTGACTTGCT 26122 4528 GTTAGGCGACGCTGACTTGC 26121 4529 GGTTAGGCGACGCTGACTTG 26120 4530 AGGTTAGGCGACGCTGACTT 26119 4531 GAGGTTAGGCGACGCTGACT 26118 4532 AGAGGTTAGGCGACGCTGAC 26117 4533 AATCGAGAAGAACCGGCTTTCGG 26161 4534 ATCGAGAAGAACCGGCTTTC 26162 4535 TCGAGAAGAACCGGCTTTCG 26163 4536 CGAGAAGAACCGGCTTTCGG 26164 4537 GAGAAGAACCGGCTTTCGGC 26165 4538 AGAAGAACCGGCTTTCGGCC 26166 4539 GAAGAACCGGCTTTCGGCCA 26167 4540 AAGAACCGGCTTTCGGCCAG 26168 4541 AGAACCGGCTTTCGGCCAGC 26169 4542 GAACCGGCTTTCGGCCAGCC 26170 4543 AACCGGCTTTCGGCCAGCCA 26171 4544 ACCGGCTTTCGGCCAGCCAG 26172 4545 CCGGCTTTCGGCCAGCCAGG 26173 4546 CGGCTTTCGGCCAGCCAGGA 26174 4547 GGCTTTCGGCCAGCCAGGAG 26175 4548 GCTTTCGGCCAGCCAGGAGT 26176 4549 CTTTCGGCCAGCCAGGAGTG 26177 4550 TTTCGGCCAGCCAGGAGTGG 26178 4551 TTCGGCCAGCCAGGAGTGGC 26179 4552 TCGGCCAGCCAGGAGTGGCC 26180 4553 CGGCCAGCCAGGAGTGGCCA 26181 4554 TAATCGAGAAGAACCGGCTT 26160 4555 GTAATCGAGAAGAACCGGCT 26159 4556 CTTTGACGCGTCCTCTCCGGGC 26295 4557 TTTGACGCGTCCTCTCCGGG 26296 4558 TTGACGCGTCCTCTCCGGGC 26297 4559 TGACGCGTCCTCTCCGGGCA 26298 4560 GACGCGTCCTCTCCGGGCAC 26299 4561 ACGCGTCCTCTCCGGGCACT 26300 4562 CGCGTCCTCTCCGGGCACTT 26301 4563 GCGTCCTCTCCGGGCACTTT 26302 4564 CGTCCTCTCCGGGCACTTTA 26303 4565 GTCCTCTCCGGGCACTTTAA 26304 4566 TCCTCTCCGGGCACTTTAAT 26305 4567 CCTCTCCGGGCACTTTAATA 26306 4568 CTCTCCGGGCACTTTAATAC 26307 4569 TCTCCGGGCACTTTAATACC 26308 4570 CTCCGGGCACTTTAATACCA 26309 4571 TCCGGGCACTTTAATACCAA 26310 4572 CCGGGCACTTTAATACCAAA 26311 4573 ACTTTGACGCGTCCTCTCCG 26294 4574 AACTTTGACGCGTCCTCTCC 26293 4575 CAACTTTGACGCGTCCTCTC 26292 4576 CCAACTTTGACGCGTCCTCT 26291 4577 TCCAACTTTGACGCGTCCTC 26290 4578 GTCCAACTTTGACGCGTCCT 26289 4579 TGTCCAACTTTGACGCGTCC 26288 4580 GTGTCCAACTTTGACGCGTC 26287 4581 AGTGTCCAACTTTGACGCGT 26286 4582 CAGTGTCCAACTTTGACGCG 26285 4583 ACAGTGTCCAACTTTGACGC 26284 4584 CACAGTGTCCAACTTTGACG 26283 4585 CGGCTCCGCCACTTGACAGCTATGTGG 26334 4586 GGCTCCGCCACTTGACAGCT 26335 4587 GCTCCGCCACTTGACAGCTA 26336 4588 CTCCGCCACTTGACAGCTAT 26337 4589 TCCGCCACTTGACAGCTATG 26338 4590 CCGCCACTTGACAGCTATGT 26339 4591 CGCCACTTGACAGCTATGTG 26340 4592 ACGGCTCCGCCACTTGACAG 26333 4593 CACGGCTCCGCCACTTGACA 26332 4594 TCACGGCTCCGCCACTTGAC 26331 4595 ATCACGGCTCCGCCACTTGA 26330 4596 AATCACGGCTCCGCCACTTG 26329 4597 AAATCACGGCTCCGCCACTT 26328 4598 CAAATCACGGCTCCGCCACT 26327 4599 CCAAATCACGGCTCCGCCAC 26326 4600 ACCAAATCACGGCTCCGCCA 26325 4601 TACCAAATCACGGCTCCGCC 26324 4602 ATACCAAATCACGGCTCCGC 26323 4603 AATACCAAATCACGGCTCCG 26322 4604 TAATACCAAATCACGGCTCC 26321 4605 AGGCGGAGATTGCGGTGAGCCGAAATCGCG 27188 4606 GGCGGAGATTGCGGTGAGCC 27189 4607 GCGGAGATTGCGGTGAGCCG 27190 4608 CGGAGATTGCGGTGAGCCGA 27191 4609 AGGCCGCCCCAACGTCCTGTCGTTCGGCGG 25618 4610 GGCCGCCCCAACGTCCTGTC 25619 4611 GCCGCCCCAACGTCCTGTCG 25620 4612 CCGCCCCAACGTCCTGTCGT 25621 4613 CGCCCCAACGTCCTGTCGTT 25622 4614 GCCCCAACGTCCTGTCGTTC 25623 4615 CCCCAACGTCCTGTCGTTCG 25624 4616 CCCAACGTCCTGTCGTTCGG 25625 4617 CCAACGTCCTGTCGTTCGGC 25626 4618 CAACGTCCTGTCGTTCGGCG 25627 4619 AACGTCCTGTCGTTCGGCGG 25628 4620 ACGTCCTGTCGTTCGGCGGC 25629 4621 CGTCCTGTCGTTCGGCGGCA 25630 4622 GTCCTGTCGTTCGGCGGCAG 25631 4623 TCCTGTCGTTCGGCGGCAGC 25632 4624 CCTGTCGTTCGGCGGCAGCT 25633 4625 CTGTCGTTCGGCGGCAGCTT 25634 4626 TGTCGTTCGGCGGCAGCTTC 25635 4627 GTCGTTCGGCGGCAGCTTCT 25636 4628 TCGTTCGGCGGCAGCTTCTC 25637 4629 CGTTCGGCGGCAGCTTCTCG 25638 4630 GTTCGGCGGCAGCTTCTCGC 25639 4631 TTCGGCGGCAGCTTCTCGCC 25640 4632 TCGGCGGCAGCTTCTCGCCC 25641 4633 CGGCGGCAGCTTCTCGCCCG 25642 4634 GGCGGCAGCTTCTCGCCCGC 25643 4635 GCGGCAGCTTCTCGCCCGCT 25644 4636 CGGCAGCTTCTCGCCCGCTC 25645 4637 GGCAGCTTCTCGCCCGCTCC 25646 4638 GCAGCTTCTCGCCCGCTCCT 25647 4639 CAGCTTCTCGCCCGCTCCTC 25648 4640 AGCTTCTCGCCCGCTCCTCC 25649 4641 GCTTCTCGCCCGCTCCTCCT 25650 4642 CTTCTCGCCCGCTCCTCCTC 25651 4643 TTCTCGCCCGCTCCTCCTCC 25652 4644 TCTCGCCCGCTCCTCCTCCC 25653 4645 CTCGCCCGCTCCTCCTCCCC 25654 4646 TCGCCCGCTCCTCCTCCCCG 25655 4647 CGCCCGCTCCTCCTCCCCGC 25656 4648 GCCCGCTCCTCCTCCCCGCG 25657 4649 CCCGCTCCTCCTCCCCGCGG 25658 4650 CCGCTCCTCCTCCCCGCGGC 25659 4651 CGCTCCTCCTCCCCGCGGCG 25660 4652 GCTCCTCCTCCCCGCGGCGG 25661 4653 CTCCTCCTCCCCGCGGCGGG 25662 4654 TCCTCCTCCCCGCGGCGGGT 25663 4655 CCTCCTCCCCGCGGCGGGTG 25664 4656 CTCCTCCCCGCGGCGGGTGA 25665 4657 TCCTCCCCGCGGCGGGTGAG 25666 4658 CCTCCCCGCGGCGGGTGAGG 25667 4659 CTCCCCGCGGCGGGTGAGGG 25668 4660 TCCCCGCGGCGGGTGAGGGA 25669 4661 CCCCGCGGCGGGTGAGGGAG 25670 4662 CCCGCGGCGGGTGAGGGAGC 25671 4663 CAGGCCGCCCCAACGTCCTG 25617 4664 CCAGGCCGCCCCAACGTCCT 25616 4665 GCCAGGCCGCCCCAACGTCC 25615 4666 AGCCAGGCCGCCCCAACGTC 25614 4667 GAGCCAGGCCGCCCCAACGT 25613 4668 GGAGCCAGGCCGCCCCAACG 25612 4669 GGGAGCCAGGCCGCCCCAAC 25611 4670 AGGGAGCCAGGCCGCCCCAA 25610 4671 GAGGGAGCCAGGCCGCCCCA 25609 4672 TGAGGGAGCCAGGCCGCCCC 25608 4673 CTGAGGGAGCCAGGCCGCCC 25607 4674 CCTGAGGGAGCCAGGCCGCC 25606 4675 ACCTGAGGGAGCCAGGCCGC 25605 4676 TACCTGAGGGAGCCAGGCCG 25604 4677 TCTCGCCCGCTCCTCCTCCCCGCGGCGGGTG 25653 4678 CTCGCCCGCTCCTCCTCCCC 25654 4679 TCGCCCGCTCCTCCTCCCCG 25655 4680 CGCCCGCTCCTCCTCCCCGC 25656 4681 GCCCGCTCCTCCTCCCCGCG 25657 4682 CCCGCTCCTCCTCCCCGCGG 25658 4683 CCGCTCCTCCTCCCCGCGGC 25659 4684 CGCTCCTCCTCCCCGCGGCG 25660 4685 GCTCCTCCTCCCCGCGGCGG 25661 4686 CTCCTCCTCCCCGCGGCGGG 25662 4687 TCCTCCTCCCCGCGGCGGGT 25663 4688 CCTCCTCCCCGCGGCGGGTG 25664 4689 CTCCTCCCCGCGGCGGGTGA 25665 4690 TCCTCCCCGCGGCGGGTGAG 25666 4691 CCTCCCCGCGGCGGGTGAGG 25667 4692 CTCCCCGCGGCGGGTGAGGG 25668 4693 TCCCCGCGGCGGGTGAGGGA 25669 4694 CCCCGCGGCGGGTGAGGGAG 25670 4695 CCCGCGGCGGGTGAGGGAGC 25671 4696 TTCTCGCCCGCTCCTCCTCC 25652 4697 CTTCTCGCCCGCTCCTCCTC 25651 4698 GCTTCTCGCCCGCTCCTCCT 25650 4699 AGCTTCTCGCCCGCTCCTCC 25649 4700 CAGCTTCTCGCCCGCTCCTC 25648 4701 GCAGCTTCTCGCCCGCTCCT 25647 4702 GGCAGCTTCTCGCCCGCTCC 25646 4703 CGGCAGCTTCTCGCCCGCTC 25645 4704 GCGGCAGCTTCTCGCCCGCT 25644 4705 GGCGGCAGCTTCTCGCCCGC 25643 4706 CGGCGGCAGCTTCTCGCCCG 25642 4707 TCGGCGGCAGCTTCTCGCCC 25641 4708 TTCGGCGGCAGCTTCTCGCC 25640 4709 GTTCGGCGGCAGCTTCTCGC 25639 4710 CGTTCGGCGGCAGCTTCTCG 25638 4711 TCGTTCGGCGGCAGCTTCTC 25637 4712 GTCGTTCGGCGGCAGCTTCT 25636 4713 TGTCGTTCGGCGGCAGCTTC 25635 4714 CTGTCGTTCGGCGGCAGCTT 25634 4715 CCTGTCGTTCGGCGGCAGCT 25633 4716 TCCTGTCGTTCGGCGGCAGC 25632 4717 GTCCTGTCGTTCGGCGGCAG 25631 4718 CGTCCTGTCGTTCGGCGGCA 25630 4719 ACGTCCTGTCGTTCGGCGGC 25629 4720 AACGTCCTGTCGTTCGGCGG 25628 4721 CAACGTCCTGTCGTTCGGCG 25627 4722 CCAACGTCCTGTCGTTCGGC 25626 4723 CCCAACGTCCTGTCGTTCGG 25625 4724 CCCCAACGTCCTGTCGTTCG 25624 4725 GCCCCAACGTCCTGTCGTTC 25623 4726 CGCCCCAACGTCCTGTCGTT 25622 4727 CCGCCCCAACGTCCTGTCGT 25621 4728 GCCGCCCCAACGTCCTGTCG 25620 4729 GGCCGCCCCAACGTCCTGTC 25619 4730 AGGCCGCCCCAACGTCCTGT 25618 4731 CAGGCCGCCCCAACGTCCTG 25617 4732 CCAGGCCGCCCCAACGTCCT 25616 4733 GCCAGGCCGCCCCAACGTCC 25615 4734 AGCCAGGCCGCCCCAACGTC 25614 4735 GAGCCAGGCCGCCCCAACGT 25613 4736 GGAGCCAGGCCGCCCCAACG 25612 4737 GGGAGCCAGGCCGCCCCAAC 25611 4738 AGGGAGCCAGGCCGCCCCAA 25610 4739 GAGGGAGCCAGGCCGCCCCA 25609 4740 TGAGGGAGCCAGGCCGCCCC 25608 4741 CTGAGGGAGCCAGGCCGCCC 25607 4742 CCTGAGGGAGCCAGGCCGCC 25606 4743 ACCTGAGGGAGCCAGGCCGC 25605 4744 TACCTGAGGGAGCCAGGCCG 25604 4745 CGGGAGGCGGTCACATTCGGCGCG 25690 4746 GGGAGGCGGTCACATTCGGC 25691 4747 GGAGGCGGTCACATTCGGCG 25692 4748 GAGGCGGTCACATTCGGCGC 25693 4749 AGGCGGTCACATTCGGCGCG 25694 4750 GGCGGTCACATTCGGCGCGT 25695 4751 GCGGTCACATTCGGCGCGTC 25696 4752 CGGTCACATTCGGCGCGTCC 25697 4753 GGTCACATTCGGCGCGTCCC 25698 4754 GTCACATTCGGCGCGTCCCC 25699 4755 TCACATTCGGCGCGTCCCCA 25700 4756 CACATTCGGCGCGTCCCCAG 25701 4757 ACATTCGGCGCGTCCCCAGC 25702 4758 CATTCGGCGCGTCCCCAGCC 25703 4759 ATTCGGCGCGTCCCCAGCCC 25704 4760 TTCGGCGCGTCCCCAGCCCA 25705 4761 TCGGCGCGTCCCCAGCCCAG 25706 4762 CGGCGCGTCCCCAGCCCAGG 25707 4763 GGCGCGTCCCCAGCCCAGGG 25708 4764 GCGCGTCCCCAGCCCAGGGG 25709 4765 CGCGTCCCCAGCCCAGGGGA 25710 4766 GCGTCCCCAGCCCAGGGGAC 25711 4767 CGTCCCCAGCCCAGGGGACG 25712 4768 GTCCCCAGCCCAGGGGACGG 25713 4769 TCCCCAGCCCAGGGGACGGA 25714 4770 CCCCAGCCCAGGGGACGGAG 25715 4771 CCCAGCCCAGGGGACGGAGC 25716 4772 CCAGCCCAGGGGACGGAGCC 25717 4773 CAGCCCAGGGGACGGAGCCC 25718 4774 AGCCCAGGGGACGGAGCCCC 25719 4775 GCCCAGGGGACGGAGCCCCG 25720 4776 CCCAGGGGACGGAGCCCCGA 25721 4777 CCAGGGGACGGAGCCCCGAG 25722 4778 CAGGGGACGGAGCCCCGAGC 25723 4779 GCGGGAGGCGGTCACATTCG 25689 4780 AGCGGGAGGCGGTCACATTC 25688 4781 GAGCGGGAGGCGGTCACATT 25687 4782 CGGAGCCCCGAGCAGCCCCCGCATCG 25730 4783 GGAGCCCCGAGCAGCCCCCG 25731 4784 GAGCCCCGAGCAGCCCCCGC 25732 4785 AGCCCCGAGCAGCCCCCGCA 25733 4786 GCCCCGAGCAGCCCCCGCAT 25734 4787 CCCCGAGCAGCCCCCGCATC 25735 4788 CCCGAGCAGCCCCCGCATCG 25736 4789 CCGAGCAGCCCCCGCATCGT 25737 4790 CGAGCAGCCCCCGCATCGTA 25738 4791 GAGCAGCCCCCGCATCGTAG 25739 4792 AGCAGCCCCCGCATCGTAGC 25740 4793 GCAGCCCCCGCATCGTAGCA 25741 4794 CAGCCCCCGCATCGTAGCAA 25742 4795 AGCCCCCGCATCGTAGCAAA 25743 4796 GCCCCCGCATCGTAGCAAAC 25744 4797 CCCCCGCATCGTAGCAAACG 25745 4798 CCCCGCATCGTAGCAAACGC 25746 4799 CCCGCATCGTAGCAAACGCG 25747 4800 CCGCATCGTAGCAAACGCGC 25748 4801 CGCATCGTAGCAAACGCGCT 25749 4802 GCATCGTAGCAAACGCGCTC 25750 4803 CATCGTAGCAAACGCGCTCC 25751 4804 ATCGTAGCAAACGCGCTCCG 25752 4805 TCGTAGCAAACGCGCTCCGC 25753 4806 CGTAGCAAACGCGCTCCGCG 25754 4807 GTAGCAAACGCGCTCCGCGC 25755 4808 TAGCAAACGCGCTCCGCGCC 25756 4809 AGCAAACGCGCTCCGCGCCT 25757 4810 GCAAACGCGCTCCGCGCCTC 25758 4811 CAAACGCGCTCCGCGCCTCA 25759 4812 AAACGCGCTCCGCGCCTCAG 25760 4813 AACGCGCTCCGCGCCTCAGG 25761 4814 ACGCGCTCCGCGCCTCAGGG 25762 4815 CGCGCTCCGCGCCTCAGGGC 25763 4816 GCGCTCCGCGCCTCAGGGCA 25764 4817 CGCTCCGCGCCTCAGGGCAC 25765 4818 GCTCCGCGCCTCAGGGCACG 25766 4819 CTCCGCGCCTCAGGGCACGC 25767 4820 TCCGCGCCTCAGGGCACGCG 25768 4821 CCGCGCCTCAGGGCACGCGC 25769 4822 CGCGCCTCAGGGCACGCGCC 25770 4823 GCGCCTCAGGGCACGCGCCC 25771 4824 CGCCTCAGGGCACGCGCCCC 25772 4825 GCCTCAGGGCACGCGCCCCA 25773 4826 CCTCAGGGCACGCGCCCCAA 25774 4827 CTCAGGGCACGCGCCCCAAA 25775 4828 TCAGGGCACGCGCCCCAAAG 25776 4829 CAGGGCACGCGCCCCAAAGC 25777 4830 AGGGCACGCGCCCCAAAGCC 25778 4831 GGGCACGCGCCCCAAAGCCC 25779 4832 GGCACGCGCCCCAAAGCCCG 25780 4833 GCACGCGCCCCAAAGCCCGG 25781 4834 CACGCGCCCCAAAGCCCGGC 25782 4835 ACGCGCCCCAAAGCCCGGCC 25783 4836 CGCGCCCCAAAGCCCGGCCA 25784 4837 GCGCCCCAAAGCCCGGCCAG 25785 4838 CGCCCCAAAGCCCGGCCAGC 25786 4839 GCCCCAAAGCCCGGCCAGCT 25787 4840 CCCCAAAGCCCGGCCAGCTG 25788 4841 CCCAAAGCCCGGCCAGCTGA 25789 4842 CCAAAGCCCGGCCAGCTGAC 25790 4843 CAAAGCCCGGCCAGCTGACC 25791 4844 AAAGCCCGGCCAGCTGACCC 25792 4845 AAGCCCGGCCAGCTGACCCT 25793 4846 AGCCCGGCCAGCTGACCCTT 25794 4847 GCCCGGCCAGCTGACCCTTT 25795 4848 CCCGGCCAGCTGACCCTTTT 25796 4849 CCGGCCAGCTGACCCTTTTC 25797 4850 CGGCCAGCTGACCCTTTTCG 25798 4851 GGCCAGCTGACCCTTTTCGG 25799 4852 GCCAGCTGACCCTTTTCGGG 25800 4853 CCAGCTGACCCTTTTCGGGG 25801 4854 CAGCTGACCCTTTTCGGGGC 25802 4855 AGCTGACCCTTTTCGGGGCC 25803 4856 GCTGACCCTTTTCGGGGCCC 25804 4857 CTGACCCTTTTCGGGGCCCA 25805 4858 TGACCCTTTTCGGGGCCCAA 25806 4859 GACCCTTTTCGGGGCCCAAA 25807 4860 ACCCTTTTCGGGGCCCAAAA 25808 4861 CCCTTTTCGGGGCCCAAAAA 25809 4862 CCTTTTCGGGGCCCAAAAAA 25810 4863 CTTTTCGGGGCCCAAAAAAG 25811 4864 TTTTCGGGGCCCAAAAAAGG 25812 4865 TTTCGGGGCCCAAAAAAGGC 25813 4866 TTCGGGGCCCAAAAAAGGCA 25814 4867 ACGGAGCCCCGAGCAGCCCC 25729 4868 GACGGAGCCCCGAGCAGCCC 25728 4869 GGACGGAGCCCCGAGCAGCC 25727 4870 GGGACGGAGCCCCGAGCAGC 25726 4871 CGCGCTCCGCGCCTCAGGGCACGCGCC 25763 4872 GCGCTCCGCGCCTCAGGGCA 25764 4873 CGCTCCGCGCCTCAGGGCAC 25765 4874 GCTCCGCGCCTCAGGGCACG 25766 4875 CTCCGCGCCTCAGGGCACGC 25767 4876 TCCGCGCCTCAGGGCACGCG 25768 4877 CCGCGCCTCAGGGCACGCGC 25769 4878 CGCGCCTCAGGGCACGCGCC 25770 4879 GCGCCTCAGGGCACGCGCCC 25771 4880 CGCCTCAGGGCACGCGCCCC 25772 4881 GCCTCAGGGCACGCGCCCCA 25773 4882 CCTCAGGGCACGCGCCCCAA 25774 4883 CTCAGGGCACGCGCCCCAAA 25775 4884 TCAGGGCACGCGCCCCAAAG 25776 4885 CAGGGCACGCGCCCCAAAGC 25777 4886 AGGGCACGCGCCCCAAAGCC 25778 4887 GGGCACGCGCCCCAAAGCCC 25779 4888 GGCACGCGCCCCAAAGCCCG 25780 4889 GCACGCGCCCCAAAGCCCGG 25781 4890 CACGCGCCCCAAAGCCCGGC 25782 4891 ACGCGCCCCAAAGCCCGGCC 25783 4892 CGCGCCCCAAAGCCCGGCCA 25784 4893 GCGCCCCAAAGCCCGGCCAG 25785 4894 CGCCCCAAAGCCCGGCCAGC 25786 4895 GCCCCAAAGCCCGGCCAGCT 25787 4896 CCCCAAAGCCCGGCCAGCTG 25788 4897 CCCAAAGCCCGGCCAGCTGA 25789 4898 CCAAAGCCCGGCCAGCTGAC 25790 4899 CAAAGCCCGGCCAGCTGACC 25791 4900 AAAGCCCGGCCAGCTGACCC 25792 4901 AAGCCCGGCCAGCTGACCCT 25793 4902 AGCCCGGCCAGCTGACCCTT 25794 4903 GCCCGGCCAGCTGACCCTTT 25795 4904 CCCGGCCAGCTGACCCTTTT 25796 4905 CCGGCCAGCTGACCCTTTTC 25797 4906 CGGCCAGCTGACCCTTTTCG 25798 4907 GGCCAGCTGACCCTTTTCGG 25799 4908 GCCAGCTGACCCTTTTCGGG 25800 4909 CCAGCTGACCCTTTTCGGGG 25801 4910 CAGCTGACCCTTTTCGGGGC 25802 4911 AGCTGACCCTTTTCGGGGCC 25803 4912 GCTGACCCTTTTCGGGGCCC 25804 4913 CTGACCCTTTTCGGGGCCCA 25805 4914 TGACCCTTTTCGGGGCCCAA 25806 4915 GACCCTTTTCGGGGCCCAAA 25807 4916 ACCCTTTTCGGGGCCCAAAA 25808 4917 CCCTTTTCGGGGCCCAAAAA 25809 4918 CCTTTTCGGGGCCCAAAAAA 25810 4919 CTTTTCGGGGCCCAAAAAAG 25811 4920 TTTTCGGGGCCCAAAAAAGG 25812 4921 TTTCGGGGCCCAAAAAAGGC 25813 4922 TTCGGGGCCCAAAAAAGGCA 25814 4923 ACGCGCTCCGCGCCTCAGGG 25762 4924 AACGCGCTCCGCGCCTCAGG 25761 4925 AAACGCGCTCCGCGCCTCAG 25760 4926 CAAACGCGCTCCGCGCCTCA 25759 4927 GCAAACGCGCTCCGCGCCTC 25758 4928 AGCAAACGCGCTCCGCGCCT 25757 4929 TAGCAAACGCGCTCCGCGCC 25756 4930 GTAGCAAACGCGCTCCGCGC 25755 4931 CGTAGCAAACGCGCTCCGCG 25754 4932 TCGTAGCAAACGCGCTCCGC 25753 4933 ATCGTAGCAAACGCGCTCCG 25752 4934 CATCGTAGCAAACGCGCTCC 25751 4935 GCATCGTAGCAAACGCGCTC 25750 4936 CGCATCGTAGCAAACGCGCT 25749 4937 CCGCATCGTAGCAAACGCGC 25748 4938 CCCGCATCGTAGCAAACGCG 25747 4939 CCCCGCATCGTAGCAAACGC 25746 4940 CCCCCGCATCGTAGCAAACG 25745 4941 GCCCCCGCATCGTAGCAAAC 25744 4942 AGCCCCCGCATCGTAGCAAA 25743 4943 CAGCCCCCGCATCGTAGCAA 25742 4944 GCAGCCCCCGCATCGTAGCA 25741 4945 AGCAGCCCCCGCATCGTAGC 25740 4946 GAGCAGCCCCCGCATCGTAG 25739 4947 CGAGCAGCCCCCGCATCGTA 25738 4948 CCGAGCAGCCCCCGCATCGT 25737 4949 CCCGAGCAGCCCCCGCATCG 25736 4950 CCCCGAGCAGCCCCCGCATC 25735 4951 GCCCCGAGCAGCCCCCGCAT 25734 4952 AGCCCCGAGCAGCCCCCGCA 25733 4953 GAGCCCCGAGCAGCCCCCGC 25732 4954 GGAGCCCCGAGCAGCCCCCG 25731 4955 CGGAGCCCCGAGCAGCCCCC 25730 4956 ACGGAGCCCCGAGCAGCCCC 25729 4957 GACGGAGCCCCGAGCAGCCC 25728 4958 GGACGGAGCCCCGAGCAGCC 25727 4959 GGGACGGAGCCCCGAGCAGC 25726 4960 AGCCGTTCCCGCCTCACAATCG 25840 4961 GCCGTTCCCGCCTCACAATC 25841 4962 CCGTTCCCGCCTCACAATCG 25842 4963 CGTTCCCGCCTCACAATCGT 25843 4964 GTTCCCGCCTCACAATCGTT 25844 4965 TTCCCGCCTCACAATCGTTT 25845 4966 TCCCGCCTCACAATCGTTTT 25846 4967 CCCGCCTCACAATCGTTTTC 25847 4968 CCGCCTCACAATCGTTTTCC 25848 4969 CGCCTCACAATCGTTTTCCT 25849 4970 GCCTCACAATCGTTTTCCTC 25850 4971 CCTCACAATCGTTTTCCTCT 25851 4972 AAGCCGTTCCCGCCTCACAA 25839 4973 AAAGCCGTTCCCGCCTCACA 25838 4974 GAAAGCCGTTCCCGCCTCAC 25837 4975 AGAAAGCCGTTCCCGCCTCA 25836 4976 CAGAAAGCCGTTCCCGCCTC 25835 4977 GCAGAAAGCCGTTCCCGCCT 25834 4978 AGCAGAAAGCCGTTCCCGCC 25833 4979 CAGCAGAAAGCCGTTCCCGC 25832 4980 GCAGCAGAAAGCCGTTCCCG 25831 4981 GGCAGCAGAAAGCCGTTCCC 25830 4982 AGGCAGCAGAAAGCCGTTCC 25829 4983 AAGGCAGCAGAAAGCCGTTC 25828 4984 CCGCCATCTAAGATGGCGGCC 25876 4985 CGCCATCTAAGATGGCGGCC 25877 4986 GCCATCTAAGATGGCGGCCC 25878 4987 CCATCTAAGATGGCGGCCCA 25879 4988 CATCTAAGATGGCGGCCCAA 25880 4989 ATCTAAGATGGCGGCCCAAG 25881 4990 TCTAAGATGGCGGCCCAAGC 25882 4991 CTAAGATGGCGGCCCAAGCG 25883 4992 TAAGATGGCGGCCCAAGCGC 25884 4993 AAGATGGCGGCCCAAGCGCC 25885 4994 AGATGGCGGCCCAAGCGCCC 25886 4995 GATGGCGGCCCAAGCGCCCG 25887 4996 ATGGCGGCCCAAGCGCCCGC 25888 4997 TGGCGGCCCAAGCGCCCGCG 25889 4998 GGCGGCCCAAGCGCCCGCGA 25890 4999 GCGGCCCAAGCGCCCGCGAT 25891 5000 CGGCCCAAGCGCCCGCGATT 25892 5001 GGCCCAAGCGCCCGCGATTA 25893 5002 GCCCAAGCGCCCGCGATTAA 25894 5003 CCCAAGCGCCCGCGATTAAG 25895 5004 CCAAGCGCCCGCGATTAAGA 25896 5005 CAAGCGCCCGCGATTAAGAC 25897 5006 AAGCGCCCGCGATTAAGACT 25898 5007 AGCGCCCGCGATTAAGACTC 25899 5008 GCGCCCGCGATTAAGACTCT 25900 5009 CGCCCGCGATTAAGACTCTC 25901 5010 GCCCGCGATTAAGACTCTCG 25902 5011 CCCGCGATTAAGACTCTCGG 25903 5012 CCGCGATTAAGACTCTCGGG 25904 5013 CGCGATTAAGACTCTCGGGC 25905 5014 GCGATTAAGACTCTCGGGCG 25906 5015 CGATTAAGACTCTCGGGCGG 25907 5016 GATTAAGACTCTCGGGCGGC 25908 5017 ATTAAGACTCTCGGGCGGCC 25909 5018 TTAAGACTCTCGGGCGGCCC 25910 5019 TAAGACTCTCGGGCGGCCCA 25911 5020 AAGACTCTCGGGCGGCCCAG 25912 5021 AGACTCTCGGGCGGCCCAGA 25913 5022 GACTCTCGGGCGGCCCAGAC 25914 5023 ACTCTCGGGCGGCCCAGACG 25915 5024 CTCTCGGGCGGCCCAGACGA 25916 5025 TCTCGGGCGGCCCAGACGAG 25917 5026 CTCGGGCGGCCCAGACGAGC 25918 5027 TCGGGCGGCCCAGACGAGCG 25919 5028 CGGGCGGCCCAGACGAGCGA 25920 5029 CCCGCCATCTAAGATGGCGG 25875 5030 TCCCGCCATCTAAGATGGCG 25874 5031 CTCCCGCCATCTAAGATGGC 25873 5032 ACTCCCGCCATCTAAGATGG 25872 5033 TACTCCCGCCATCTAAGATG 25871 5034 TTACTCCCGCCATCTAAGAT 25870 5035 CTTACTCCCGCCATCTAAGA 25869 5036 TCTTACTCCCGCCATCTAAG 25868 5037 CTCTTACTCCCGCCATCTAA 25867 5038 CCTCTTACTCCCGCCATCTA 25866 5039 TCCTCTTACTCCCGCCATCT 25865 5040 TTCCTCTTACTCCCGCCATC 25864 5041 TTTCCTCTTACTCCCGCCAT 25863 5042 TTTTCCTCTTACTCCCGCCA 25862 5043 GTTTTCCTCTTACTCCCGCC 25861 5044 CGTTTTCCTCTTACTCCCGC 25860 5045 TCGTTTTCCTCTTACTCCCG 25859 5046 ATCGTTTTCCTCTTACTCCC 25858 5047 CGGGCGGCCCAGACGAGCGAGCCCTCG 25920 5048 TCGGGCGGCCCAGACGAGCG 25919 5049 CTCGGGCGGCCCAGACGAGC 25918 5050 TCTCGGGCGGCCCAGACGAG 25917 5051 CTCTCGGGCGGCCCAGACGA 25916 5052 ACTCTCGGGCGGCCCAGACG 25915 5053 GACTCTCGGGCGGCCCAGAC 25914 5054 AGACTCTCGGGCGGCCCAGA 25913 5055 AAGACTCTCGGGCGGCCCAG 25912 5056 TAAGACTCTCGGGCGGCCCA 25911 5057 TTAAGACTCTCGGGCGGCCC 25910 5058 ATTAAGACTCTCGGGCGGCC 25909 5059 GATTAAGACTCTCGGGCGGC 25908 5060 CGATTAAGACTCTCGGGCGG 25907 5061 GCGATTAAGACTCTCGGGCG 25906 5062 CGCGATTAAGACTCTCGGGC 25905 5063 CCGCGATTAAGACTCTCGGG 25904 5064 CCCGCGATTAAGACTCTCGG 25903 5065 GCCCGCGATTAAGACTCTCG 25902 5066 CGCCCGCGATTAAGACTCTC 25901 5067 GCGCCCGCGATTAAGACTCT 25900 5068 AGCGCCCGCGATTAAGACTC 25899 5069 AAGCGCCCGCGATTAAGACT 25898 5070 CAAGCGCCCGCGATTAAGAC 25897 5071 CCAAGCGCCCGCGATTAAGA 25896 5072 CCCAAGCGCCCGCGATTAAG 25895 5073 GCCCAAGCGCCCGCGATTAA 25894 5074 GGCCCAAGCGCCCGCGATTA 25893 5075 CGGCCCAAGCGCCCGCGATT 25892 5076 GCGGCCCAAGCGCCCGCGAT 25891 5077 GGCGGCCCAAGCGCCCGCGA 25890 5078 TGGCGGCCCAAGCGCCCGCG 25889 5079 ATGGCGGCCCAAGCGCCCGC 25888 5080 GATGGCGGCCCAAGCGCCCG 25887 5081 AGATGGCGGCCCAAGCGCCC 25886 5082 AAGATGGCGGCCCAAGCGCC 25885 5083 TAAGATGGCGGCCCAAGCGC 25884 5084 CTAAGATGGCGGCCCAAGCG 25883 5085 TCTAAGATGGCGGCCCAAGC 25882 5086 ATCTAAGATGGCGGCCCAAG 25881 5087 CATCTAAGATGGCGGCCCAA 25880 5088 CCATCTAAGATGGCGGCCCA 25879 5089 GCCATCTAAGATGGCGGCCC 25878 5090 CGCCATCTAAGATGGCGGCC 25877 5091 CCGCCATCTAAGATGGCGGC 25876 5092 CCCGCCATCTAAGATGGCGG 25875 5093 TCCCGCCATCTAAGATGGCG 25874 5094 CTCCCGCCATCTAAGATGGC 25873 5095 ACTCCCGCCATCTAAGATGG 25872 5096 TACTCCCGCCATCTAAGATG 25871 5097 TTACTCCCGCCATCTAAGAT 25870 5098 CTTACTCCCGCCATCTAAGA 25869 5099 TCTTACTCCCGCCATCTAAG 25868 5100 CTCTTACTCCCGCCATCTAA 25867 5101 CCTCTTACTCCCGCCATCTA 25866 5102 TCCTCTTACTCCCGCCATCT 25865 5103 TTCCTCTTACTCCCGCCATC 25864 5104 TTTCCTCTTACTCCCGCCAT 25863 5105 TTTTCCTCTTACTCCCGCCA 25862 5106 GTTTTCCTCTTACTCCCGCC 25861 5107 CGTTTTCCTCTTACTCCCGC 25860 5108 TCGTTTTCCTCTTACTCCCG 25859 5109 ATCGTTTTCCTCTTACTCCC 25858 5110 CGTCCTCCCGACCTGCGACGCCACCGGC 25957 5111 GTCCTCCCGACCTGCGACGC 25958 5112 TCCTCCCGACCTGCGACGCC 25959 5113 CCTCCCGACCTGCGACGCCA 25960 5114 CTCCCGACCTGCGACGCCAC 25961 5115 TCCCGACCTGCGACGCCACC 25962 5116 CCCGACCTGCGACGCCACCG 25963 5117 CCGACCTGCGACGCCACCGG 25964 5118 CGACCTGCGACGCCACCGGC 25965 5119 GACCTGCGACGCCACCGGCT 25966 5120 ACCTGCGACGCCACCGGCTC 25967 5121 CCTGCGACGCCACCGGCTCT 25968 5122 CTGCGACGCCACCGGCTCTC 25969 5123 TGCGACGCCACCGGCTCTCC 25970 5124 GCGACGCCACCGGCTCTCCG 25971 5125 CGACGCCACCGGCTCTCCGA 25972 5126 GACGCCACCGGCTCTCCGAT 25973 5127 ACGCCACCGGCTCTCCGATT 25974 5128 CGCCACCGGCTCTCCGATTC 25975 5129 GCCACCGGCTCTCCGATTCT 25976 5130 CCACCGGCTCTCCGATTCTG 25977 5131 CACCGGCTCTCCGATTCTGC 25978 5132 ACCGGCTCTCCGATTCTGCG 25979 5133 CCGGCTCTCCGATTCTGCGC 25980 5134 CGGCTCTCCGATTCTGCGCG 25981 5135 GGCTCTCCGATTCTGCGCGA 25982 5136 GCTCTCCGATTCTGCGCGAG 25983 5137 CTCTCCGATTCTGCGCGAGC 25984 5138 TCTCCGATTCTGCGCGAGCC 25985 5139 CTCCGATTCTGCGCGAGCCC 25986 5140 TCCGATTCTGCGCGAGCCCT 25987 5141 CCGATTCTGCGCGAGCCCTA 25988 5142 CGATTCTGCGCGAGCCCTAC 25989 5143 GATTCTGCGCGAGCCCTACT 25990 5144 ATTCTGCGCGAGCCCTACTG 25991 5145 TTCTGCGCGAGCCCTACTGG 25992 5146 TCTGCGCGAGCCCTACTGGC 25993 5147 CTGCGCGAGCCCTACTGGCA 25994 5148 TGCGCGAGCCCTACTGGCAG 25995 5149 GCGCGAGCCCTACTGGCAGT 25996 5150 CGCGAGCCCTACTGGCAGTC 25997 5151 GCGAGCCCTACTGGCAGTCG 25998 5152 CGAGCCCTACTGGCAGTCGA 25999 5153 GAGCCCTACTGGCAGTCGAC 26000 5154 AGCCCTACTGGCAGTCGACT 26001 5155 GCCCTACTGGCAGTCGACTT 26002 5156 CCCTACTGGCAGTCGACTTC 26003 5157 CCTACTGGCAGTCGACTTCT 26004 5158 CTACTGGCAGTCGACTTCTA 26005 5159 TACTGGCAGTCGACTTCTAA 26006 5160 ACTGGCAGTCGACTTCTAAC 26007 5161 CTGGCAGTCGACTTCTAACT 26008 5162 TGGCAGTCGACTTCTAACTT 26009 5163 GGCAGTCGACTTCTAACTTG 26010 5164 GCAGTCGACTTCTAACTTGG 26011 5165 CAGTCGACTTCTAACTTGGC 26012 5166 AGTCGACTTCTAACTTGGCT 26013 5167 GTCGACTTCTAACTTGGCTC 26014 5168 TCGACTTCTAACTTGGCTCG 26015 5169 CGACTTCTAACTTGGCTCGG 26016 5170 GACTTCTAACTTGGCTCGGG 26017 5171 ACTTCTAACTTGGCTCGGGC 26018 5172 CTTCTAACTTGGCTCGGGCA 26019 5173 TTCTAACTTGGCTCGGGCAT 26020 5174 TCTAACTTGGCTCGGGCATC 26021 5175 CTAACTTGGCTCGGGCATCC 26022 5176 TAACTTGGCTCGGGCATCCA 26023 5177 AACTTGGCTCGGGCATCCAT 26024 5178 ACTTGGCTCGGGCATCCATC 26025 5179 CTTGGCTCGGGCATCCATCG 26026 5180 TTGGCTCGGGCATCCATCGC 26027 5181 TGGCTCGGGCATCCATCGCT 26028 5182 GGCTCGGGCATCCATCGCTC 26029 5183 GCTCGGGCATCCATCGCTCT 26030 5184 CTCGGGCATCCATCGCTCTG 26031 5185 TCGGGCATCCATCGCTCTGG 26032 5186 CGGGCATCCATCGCTCTGGC 26033 5187 GGGCATCCATCGCTCTGGCC 26034 5188 GGCATCCATCGCTCTGGCCT 26035 5189 GCATCCATCGCTCTGGCCTG 26036 5190 CATCCATCGCTCTGGCCTGA 26037 5191 ATCCATCGCTCTGGCCTGAA 26038 5192 TCCATCGCTCTGGCCTGAAC 26039 5193 CCATCGCTCTGGCCTGAACT 26040 5194 CATCGCTCTGGCCTGAACTC 26041 5195 ATCGCTCTGGCCTGAACTCA 26042 5196 TCGCTCTGGCCTGAACTCAG 26043 5197 CGCTCTGGCCTGAACTCAGG 26044 5198 TCGTCCTCCCGACCTGCGAC 25956 5199 CTCGTCCTCCCGACCTGCGA 25955 5200 GCTCGTCCTCCCGACCTGCG 25954 5201 TGCTCGTCCTCCCGACCTGC 25953 5202 GTGCTCGTCCTCCCGACCTG 25952 5203 GGTGCTCGTCCTCCCGACCT 25951 5204 CGGTGCTCGTCCTCCCGACC 25950 5205 TCGGTGCTCGTCCTCCCGAC 25949 5206 CTCGGTGCTCGTCCTCCCGA 25948 5207 ACTCGGTGCTCGTCCTCCCG 25947 5208 GACTCGGTGCTCGTCCTCCC 25946 5209 CGACTCGGTGCTCGTCCTCC 25945 5210 TCGACTCGGTGCTCGTCCTC 25944 5211 CTCGACTCGGTGCTCGTCCT 25943 5212 CCTCGACTCGGTGCTCGTCC 25942 5213 CCCTCGACTCGGTGCTCGTC 25941 5214 GCCCTCGACTCGGTGCTCGT 25940 5215 AGCCCTCGACTCGGTGCTCG 25939 5216 GAGCCCTCGACTCGGTGCTC 25938 5217 CGAGCCCTCGACTCGGTGCT 25937 5218 GCGAGCCCTCGACTCGGTGC 25936 5219 AGCGAGCCCTCGACTCGGTG 25935 5220 GAGCGAGCCCTCGACTCGGT 25934 5221 CGAGCGAGCCCTCGACTCGG 25933 5222 ACGAGCGAGCCCTCGACTCG 25932 5223 GACGAGCGAGCCCTCGACTC 25931 5224 AGACGAGCGAGCCCTCGACT 25930 5225 CAGACGAGCGAGCCCTCGAC 25929 5226 CCAGACGAGCGAGCCCTCGA 25928 5227 CCCAGACGAGCGAGCCCTCG 25927 5228 GCCCAGACGAGCGAGCCCTC 25926 5229 GGCCCAGACGAGCGAGCCCT 25925 5230 CGGCCCAGACGAGCGAGCCC 25924 5231 GCGGCCCAGACGAGCGAGCC 25923 5232 GGCGGCCCAGACGAGCGAGC 25922

Hot Zones (Relative upstream location to gene start site) 25500-27500

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11966) AATACAGTCTTCCCCCACAGTTGAATATAGAATAAAATCTATTGCAAGCT GGGTGCAGGGGCACAAGTGTGGCAGGAGTGCTTGAGCCTAGGAGTTCAAG ACCAGCCTGGGCAACATAGTGAGACCTCATCTCAATTGAAAATATATATC TATATAAAAAATAAAATTTATTACAGTTCATCTTGCTGGAAAACAAAATA CTGTTTTTGTAATTAAAATTTTTTTTTTAAATTTAGAAATGGGGTCTTGC TGTGTTGACCAGGCTGGTCTTGAACTCTTGGCCTCAAGCTGTCCTCCCAT CTGGGCCTCCCAAAGTGCTGGGATTACAGGTGTGAACAACTGCGCCCGGC TGACAAAGTATTTTTTAAAGATGTACCACTAAATGGAGATTTGATTCACA TTTGATAGTTTTTGACAGGTCTTTTCTATTTAAAAACATTACTGTTTTTG TAGCATTATTCTGGCTTTTCCCTTAATTTAGTAAATATTTGAGTGCCTTT GTATTCCAGATACTGAGCAAGATTGGCAGGGTTCTGCCCTTATGGAGCAG AAGGAAGGTAGGGGGACTGACTAAAACTTGAAAACTGTCTAACATAAGTA CCATGCAGAAAATGAAACAGTATTAATTGGCAGAAGGAGAGCAGGCTATT TTGGCTAGTGTGGTTAGGGAAAGCCTCTCTAAAGAGATGTCTCTTGGGTG GAGACAAGATGTGAAAAAACCAGCTTGCCTGTTTTTGGGGTTTCAGCCTT GCAGGTGAAGAGAAACACGAAGTTCAGAAGTCTTGAGGCACAAAGTCTGG CATGTTACGAAAGAAGGCCTTTAGACGCCTTGTCAGGGAGTTTAGATTTT ATTCTGAGTTTTAAAACGGGAGTGACACAATGAGTTGCATTTTAAGCCTG TTCAGGCTGTTACATGGATTATTAGGAGCTGTATCATTTCAGGCTAGTGA GATGCTCAGATGAGTCTGCCTTCTGTCTCTTCCGTCATCTATTTCTCTCT TATCTGGTCTTAAGCTCCTCCATCTTTTCCTTTTTAGTTGGAAAAAAACT CAAAGATCTAGAAAAAAGAGGAGCTGTATGTACTCCTAAAAAGGGACCTC ATAGTAACCTGGGGATAGAGTTATGTAGGAGTGAGTCAGGGCTCAGGTTG AGGCTTTAGAGGCAGGAGGCAGCGAGATCTTGTTCTGTCATCCCCTCTTA CAGAAATAAAATATGCCGATAAAAGTTTATAGTGTAATAGTAAAATATAA AAACAAAAAGTAAGTAATGTAGAAAATAAAAACCCTTCACAGTCCTGCTG AAATGATTACTGTTAACACTTTAATTCTAGAGTTCCCCATCCATTTATTT ATTTCTAGATTTCCCTCTTTGTAGATTAATATTAAAGGGTTCAGACTTGT TCATTTTTTGTTGTCTTGGATATCTTTTCCCACCTCTGTATATATGGATC TACTTTATTTATCACGTGGATATTAACATGGTTTATTTAATTCCCTATTG TTAGGTATTTGGTCTTTACCACAGTTTTTCAAGGGTATGAATAGTGCTGC AAGGAATATGCTTACACATGTTTTTATACACTTGTCTTAGGCTTCTGTAG GACAAATTTCTGGAGTAGAATACTAGGTCATTCTTTAAGAACATTTCAAA CTTTTAATAGATATTACCGTATTCTTTCCCAAAAAGAATGTACAAAGACT GTATGAGAATAACTCCATGTTGTGATCTTAAGTTGTCTCTAAACCTCTTT GGTTTTCTTAGCTGTCATCTAAGAATACTAAGTATCTAACCTCCCTCTTG ATTTGGGCATGTGATGTGATTTAGCATATAGTGGATATTCAGTTAGAAAC TTTTGGTTGAAAACAAGGTTTGGATTCTGTGGTCTTTAATTCTAGGCCAT TTCAGCTCTGACTAAAATGATTTGAGTGTTAGTGTTATATATGGGAAGGT AAGGGCTATGGAGTCAGTGCAGCCCAGTTCAGAATCCCAGTTTGCCACTT ACAAGCTGTGTGTGTGAGAATTTTCTCAACTGTAAAATGGGGACATAATT CCTACCTAGAGTAATACTGTAAGTATTAAGGTGGATAATGATTGGAATGT ATGCTGTGTATCCTGCCTCATAATAGTAAGCTTTTAGTAAATGGTAGCTA CTGTTAATAATAAAACAAGTTTCTGAAGGAGGAAGGCTTGAAAAGATGGG ATTCCTTATCAACCTCAAAGTTTTCTAAAGGAGGAAACCCTACCCCCCTT ACTTCTGCATGGTTTCTGACCATGAACTGAACTCTGAACTCTGAATGAAC TGAACTCTGAACTCTGAATGAACTGAACTCTGAACTCTGAATGTTATGGT AGAAAATTCATGGACTTTAAATTTAAACAGATAAAGAATCTGGTTATTTT ACCCACTGCTGGGGTGTTCTTGGGCAAGTAGCATGACTTCTGTGTCCAAA AAAGAAAGGGTTTGCAGTGACTGAACCTGTAATCCCAGTACTTTGGGAGG CTAAGGAGAGTGGATTGCCTGAGCTCAGGAGTTCAAGACCAGCCTGGGCA ACATAGTGAGAGCCTTTCTCAACAAAAAAAACTGTTCTTAAAAATTAGCT GGGCATGGTGATGCACGTCTGTGGTCCCAGCTATGTGGGAAGCTGAGGTA GGAGAATCATTTGAGCCTGGAAAATTGAAGCTGCAGTGAGCTGTGATCAT GTCACTGCACCCCAGCCTGGGCAACAGAGCAAGACCCTGTCTCAGAAAAT AAATTAATTAAAAAGAAAGTGTGGATGGAGGAAGGGATTAAAAATCTGGC TGGGCACGGTGGCTCATGCCTGTAATCCCAGGCGTGATTTGGGAGGCCGA GGCGGACAGATCACGAGGTCAAGAGATTGAGACCATCCTGGCCAACATGG CCAACCCCATCTCTACTAAAAATACAAAAATCAGTCGGGCGTGGTGGTGC ATGCCTGTAATCCCGGCTACTCGGGAGGCTGAGGCAGGAGAATCGCTTGA ACCTGGGAGGTTCAGTGAGCCAAGATCGCGCCACTACACTCCAGCCTGGC AATAGAGTGAGACTCTGTCTCAAAAGAAAAGAAAAGAAAAGAAAATCTTT GGGGTTCTTACACAAATTAAATGAGATAATTTATTATTATTATTTTTTTT GAGATGGAGTCTTGCTCTGTCCCCCAGGCTGGAGTGCAGTGGTGCGATCT CAGCTCACCGCAAGCTCTGCCTCCCGGGTTCACGCCATTCCCCTGCCTCA GCCTCCTGAGTAGCTGGGACTACAGGCGCCCGCCACCATGCCTGGCTAAT TTTTTGTATTTTTAGTAGAGACAGGGTATCCCTGTGTTAGCTAGGATGGT CTCGATCTCCTGACCTTGTGATCCGCCCATCTCGGCCTCCCAAAGTGCTG GGATTACAGGTATGAGCCACCATGCCCGGCTTGAGATAATTTATAAAGTG CCTAAAATACATCCTAGAAATATTAGTTTTTCTTCCTTGAAGTCATAAAT TATGGCTTACACTTTTTTTCAGGTATTTCTCATAGTACTAATGTGTTGCT CACACTCAAGGGTAGTAGTTGCTTAGGAAGAAGAGAAATGTAGTTGAAAA AGTAATAGACTAGAAGTCTTGAGACCTGGGCTCATGTTCCAAGTTGGCTT TTTTTTTTTTTTTTGGGAGATGGAGTCTCGCTCTTGTCCCCCAGCCTGGA GTGCAATGACACGATATCGACTCACTGCAACCTCCACCTCCTGGGTTCAA GTGATTTCTCCTGCCTCAGCCTCCCTAGTAGCTGGGATGACAGACACCCA CCACCATGCCTGGCTAATTTTTGTATTTTAAGTAGTGACAGCATTTTACC ATGTTAGCCAGGCTGGTCTTGAACTCCTGGCCTCAAGTGATGCGCTGGCC TCGGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCACTGTGCCTGGTC CCTTGCTAAATGTTTTGTTTTGTTTTGTTTTGTTTTTGAGGTGGAGTCTT GCTCTGTCACCCAGGCTGGAGTGCGGTGGCATGATCTCCGCTCACTGCAA GCTCCGCCTCCCAGGTTCCCGCCATTCTCCTGCCTCAGCCTCCCGAGTAG CTGGGACTACAGGCGCCCGCCACCACGCCCGGCTAATTTTTTGTATTTTT AGTAGAGATGGGGTTTCACCGTGTTAGCCAGGATGGTCTCCATCTCCTGA CCTCGTGATGCACCCACCTCGGCCTCCCAAAGTGCTGGGATTACAGGCGT GAGCCACCGTGCCCCGCAGTTGCTTGCTAAATCTTTTAACTGCTGGTCCC ATTTTCCTCATCTATGAAATATTTAATGGAAGTGTACTATTAAAGAAACT TTTCTTTGCTGATGAATGCAGGAGGTATCATTAAAAACCCACATAGTGCT ATTTTCATAATTACTCTTTATGTATTGTGTTCTTGGGTTGAATACTTTTG TTCTAGAGTTACAATTATTTGTGTTTCTTACCAGGTTTAAGAATTGTTTA AGCTGCATCA ATG

18) Beta catenin. Proto-oncogene protein Wnt-1 is a protein that in humans is encoded by the WNT1 gene (Van Ooyen et. al, 1986; Nat. Genet. 28 (3): 261-5 and Aarheden et al., 1988; Cytogenet Cell Genet 47 (1-2): 86-87). The WNT gene family consists of structurally related genes that encode secreted signaling proteins that are implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. Wnt-1 t is conserved in evolution with the protein encoded by this gene having 98% identity to the mouse Wnt1 protein at the amino acid level.

Beta-catenin (or β-catenin) is a protein that in humans is encoded by the CTNNB1 gene. β-catenin is a subunit of the cadherin protein complex and acts as an intracellular signal transducer in the Wnt signaling pathway (McDonald et al, 2009; Developmental Cell 17 (1): 9-26). Recent evidence suggests that β-catenin plays an important role in various aspects of liver biology including liver development (both embryonic and postnatal), liver regeneration following partial hepatectomy, HGF-induced hepatomegaly, liver zonation, and pathogenesis of liver cancer (Thompson and Monga, 2007; Hepatology 45 (5): 1298-305). The gene that codes for β-catenin can function as an oncogene. An increase in β-catenin production has been noted in those people with basal cell carcinoma and leads to the increase in proliferation of related tumors (Saldanha et al, 2004; Cancer Epidemiol. Biomarkers Prev. 17 (8): 2101-8. Mutations in this gene are a cause of colorectal cancer (CRC), pilomatrixoma (PTR), medulloblastoma (MDB), and ovarian cancer. Also, β-catenin binds to the product of the APC gene, which is mutated in adenomatous polyposis of the colon (reviewed in Wang et al, 2008; Cancer Epidemiol. Biomarkers Prev. 17 (8): 2101-8).

Protein: Beta-catenin Gene: CTNNB1 (Homo sapiens, chromosome 3, 41240942-41281939 [NCBI Reference Sequence: NC_(—)000003.11]; start site location: 41265560; strand: positive)

Gene Identification GeneID 1499 HGNC 2514 HPRD — MIM 116806

Targeted Sequences Relative upstream location to gene Sequence Designed start ID No: ID Sequence (5′-3′) site 5233 BC1 CGCATATTACTGGGTAAACTCTGTG 1411 5234 CACGCTGGATTTTCAAAACAGTTG 5

Target Shift Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 5233 CGCATATTACTGGGTAAACTCTGTG 1411 5234 CACGCTGGATTTTCAAAACAGTTG 5 11987 CCACGCTGGATTTTCAAAAC 4

Hot Zones (Relative upstream location to gene start site)  1-250 1400-1500

Examples

In FIG. 35, In MCF7 (human mammary breast cell line), BC1 (191) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated control values. The β-catenin sequence BC1 fits the independent and dependent DNAi motif claims.

The secondary structure for BC1 (191) is shown in FIG. 36.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11967) ACCCTGTAGGATGGGCGGGTGATGGTATGTATGTTAGATGTGTGGACATA TCTATTAAAAGTTGTGTCAGATAACAGCTGGTGCTGACAAGCCCTTGGTA AGATGGCAGCATGTTCAATATGTTCTGTGAAAATTATCTCAGTTTATGAT CTGTCAGTATTGTGGAGCTATGCATGAAAGGACTTAAAATTCTTACCCTT AAACTCAGTAACAGTGTTTCTAGAACTTCTGGTGATATGGGAAATTAAGA GAATTATTTATATGCAAAGGTGTTTATTGCAGCATTGTTGGAATAATAGA CAAAATGGGGAAGAACAAGCTCAGAATGGAGGAGGTAGCTTATAGTATAG ACATACGATACAATCCAGATGATAATATTTTATAATAGTCTTCACAAGGA ATTTTATATTTTTATTTTTAAAAATACATAGCAGTGAGTTTAATATACCA AACATACCAAAATGTCATCATTTACTGTGTGGTGGACTCATATGATGGAG ATGATAAATAAAAATATTAATTTATTTGAGGCATATATTTATGGCTGAGG AAGGAAGACAGTTATGAAGAACAGCTCATTCTGGAAACATACTAATTTTT CCCAGCCATAAAGAGATTTCCTATTTCTTTTTTTTTTCCATTTACCTTCT GTTTCCTACCTGAGAAGATTTCATACTTCTAATAACCATTTGTGTACCTA TTTAAAGACAGTACCAAAGGCATACATTTTAGTGTTTGGAGGACCAAGGG TCATTTGATGTTTGATGCTTATTGACTATTCGAGGATGACAAGACACCTT GAGAACACACACACCCACACCCACACCCACACCCTCACCCACCCACCCCA CCCCCCTCCCCGAAGAAAGCTGTGAAGGAAGAAAGCAGAAAAGAACCTGG AGTGAGTTGTAACTTAAAATGTTAGTGTTGCATGAAGTGTGTTAAAACAG GAAGATTTGAGGAAATTGCATACATTTTCTAGATGGCAAAGTATTACTGG TGACAGTTAATGAAAATGCATATGCATGTGTTTTTAGATTTACAAATTTT ACTAAGAACTTTTTAAAAATCCCTGAAGGTGTATCAAAAGTTTATCATGC TTATGAAATAGAGTAGCACTTTCTAACTTTAAAACGGGGAATAATTCTTT GGATCTTGATTATTGGAAAAGTGAATTATGAATTGCTAGTATAAAACTGT GGTTTTAAAATATGTCTGCTTTATATTTTTATGTAGCAGATTTACTCCTA GTTAATAATACTCAAACTTACTGAAAACTAAGGTAATTAAGATAATTCTG TCCTGATGGGAAGAGGAAAAATAACTTCAGTGTGAAATCTATTATATATT AGTTGTGGCAAGATTTCTCCCATTGACTTTGACTGGAGACATTTATAGGG TTAAAATCGGAAATAGCACGGTGAATTTTGAAGTATCCTTGTAGTTGGAA AGAGTATTATGTTCATATTGCCAAAAAAAAGATGCATGGATGCATTAGAC TGGATGGAAAATACATGAGAAGTTGGCTAGCCCCCTCTTTGTCAAAACAT CACTTGGTGGTGATAAAGCTGTTGGAAAACACAGCATTCTAATGTAGTCT GTAGTTTAATGATAATCTGTGTCTTGAAACATTTAGCGTAGTACTTATAC AAACCTAGATGGCATAGTGTACTGCATGCCTAGCCTATATAGTATAGCCT GTTGCTTCTAGGGTGTAAAGCTGTATAGCGTGTTACTATAGGCAGTTGAA ACAGTGGTATTTATGTATCCTTTTTTTTTTTTTTAAATTCTTTTAAGAGA CAGGGTCTTGCTCTGTTGCCCAGGCTGGATGCATTGGTGTGATCATAGCT CACTATAACCTTGAACTCCTAAGTGATCCTCTTTGCCTCAGCCTCCCCAG TGGCTAGGACTACAGGCACATACTACCACACCTGGCTAATTTTTAACATT TTTTTGTAGAGATGGAATTTCGCTGTGTTGTCCAGGCTGGTCTTGGAACT CTTGTGCTGCAGCAATCCACCCGCCTCCCAAAGTGTTAGAATTACAAGCC ACTTCGCCTGGCTTGTTTACCTAAACATAGAAAAGATCCAGTAAAAATAC AGAATTAAAATCTTGTGGGGCCACTGTAGCATATGTAGTCCATCTTGACT GAAATGTCCTTATGCAGTGCATGATTGTACTTCATAATTTTTAAGCACTC CTCCCTCTTGATTGGTACTTAGTGGATTTTATCATTTTTGTTTCTTCATA ATTCTTTCTGAAATGTCTACTGGTTGGACCTTTGATCTCCTGAATTGATC GTGATTTCTTCTGTTGTATTTTTTGTCTTTGTCATTTTTTTGTACTCTAG GCAGTTTTCTCAATTTTAGTTTCTATTCAACTTTTTGTTTTTATTTATTC TCTCCAGTATTTATGGAGATACTAAATTGAAGTGTTCTGTTTCTCTCTCC ACCCTATCCCTAGTTTCAAGTTTTATCTCAGTTTCTATGGAGTCAGTTTT TTCGTTGCTTTAAAAAAAAATTTTCCTGAAGTGATTGGTAAGTTTTGGCT AATTGGGAGCACTAGAATTGGGCCCTTAATGGTTGGCAGGGTGTGGTGGA GGAGAGACAGCCCTTAGTCCAAAGGCTCAGGCCAGAAAAAGAAAGAGGAA GGCTTTCCTTTTCCTTTCCGGAGCAGGGTTCTGCCCTAGGTCTTGCTTGG CAGTCTATTTGATTTCTTTAGCAGTTAATGCTCAGTTTTTTGGCATATGT GGATCTGCCTCCAGAGCAGGTACAAGGTGAGTGAGTCTATGCTGTTACCT AATTAGATCCCCATTTCTACCCTTTGTTTTTACTTCTCTATCTACTGATA GGTTTTTACCCTCCTTCACCTCATAGGGTTGCAGTGAAGAGCAAGATGAA TTTTTATTTATGTTGCATAAATTTTAAAAGCTAAAAAATATATATGTAAT GTTGGGAAGTCCCAGTGTACAAATGGCTATTGTAAATTTGGAACATGAAC TTGCTTTTTTCCATTGTAAAAATGAAATCATTATAAATTGCGGTCAAGTT ACTAGGTCAGCCCACACAGAGTTTACCCAGTAATATGCGTAAATGTTTTG CCTTTGCATCAACAACAAGGAAAAACAGTACTATAAAAAAATGTTCCTGG AAGCCGGATGTATCAAAGCACTTCTGAAATAGCTATATAGCCTATAGACA TGACCAGTTGGTTTCTGAGTCTGTTGACATTGGCCAAAGGAGAAGCTCAG TGTAGAACATGTTTGGAGTCTCCTTTTGCAGAAATACATTGGAGGCTGGA GTGGGGAACCAATTTTTCAGAAAGGTGGTGAAGTAGTTACATAGCCACTC TTTTAAAGACAGTCAAAAGATAGAAACTAAGGCCAGGTGTTGGCTCACAT CTGAGATAGGAAAATCACTTGAACCTGGGAGGCGGAGGTTGCAGTGAGCC CAGTATGCACCTCTGCACTCCAGCCTGGTTTGGCAAGAGACCAAAACTCT GTCTCAAAAAAAAACAAAACATAGTTCACACTTAAATATTTTATTCCATA TCTTTACATACCCAATATGTTAATTTATAGTTCAAGATGAACTTGTTTGG GACAGATTTTGTAATAAAGGAAATCGTGTTATTAGAAATATCTAGAGGCC ATGAGCCCTTAAACTGTTCTAATTTGCAAGTAGTTCCCTGTGTGATGCAG TTTTTTTCAATATTGCACAATAAAGGCAAAATACGGACAAATTAGATGAT AAGATTTATATAAATTTTTAAAATATTGATCAAAATATGTATCCATATTG GTAATATTTGTATTTATAATAAATCATTGCTGTAAATTTGAACTTAGAAA AATTTTACTAATAAAGGTGCTTTTGTGTTGCAAACTTTCATTTGAAAAGT AATTTTTCTTTGTACCAAAAAATCTAAAATTCGCTATTCTAGTCACCAAA ATTTGCTTTATGAAAAATAATTTTTGATGGCACTATATCAGAAAACAACT TGTTAAAGAAAATGTGGAGTTTTTAAAATCCCACTGTACCTCTGTTATCC AAAGGGGATCTGTGAATTTTTCTGTGAAAGGTTAAAAAAGGAGAGACCTT TAGGAATTCAGAGAGCAGCTGATTTTTGAATAGTGTTTTCCCCTCCCTGG CTTTTATTATTACAACTCTGTGCTTTTTCATCACCATCCTGAATATCTAT AATTAATATTTATACTATTAATAAAAAGACATTTTTGGTAAGGAGGAGTT TTCACTGAAGTTCAGCAGTGATGGAGCTGTGGTTGAGGTGTCTGGAGGAG ACCATGAGGTCTGCGTTTCACTAACCTGGTAAAAGAGGATATGGGTTTTT TTTGTGGGTGTAATAGTGACATTTAACAGGTATCCCAGTGACTTAGGAGT ATTAATCAAGCTAAATTTAAATCCTAATGACTTTTGATTAACTTTTTTTA GGGTATTTGAAGTATACCATACAACTGTTTTGAAAATCCAGCGTGGACA ATG

19) PCSK9. Proprotein convertase subtilisin/kexin type 9, also known as PCSK9, is an enzyme that in humans is encoded by the PCSK9 gene. This gene encodes a proprotein convertase belonging to the proteinase K subfamily of the secretory subtilase family. The encoded protein is synthesized as a soluble zymogen that undergoes autocatalytic intramolecular processing in the endoplasmic reticulum. This protein plays a major regulatory role in cholesterol homeostasis. PCSK9 binds to the epidermal growth factor-like repeat A (EGF-A) domain of the low-density lipoprotein receptor (LDLR), inducing LDLR degradation. Reduced LDLR levels result in decreased metabolism of low-density lipoproteins, which could lead to hypercholesterolemia. Variants of PCSK9 can reduce or increase circulating cholesterol. LDL cholesterol is removed from the blood when it binds to LDL receptors on the surface of liver cells, and is taken inside the cells. When PCSK9 binds to the LDL receptor, the receptor is destroyed along with the LDL. But if PCSK9 does not bind, the receptor can return to the surface of the cell and remove more cholesterol (reviewed in Akram et al, 2010 Arterioscler Thromb Vasc Biol.; 30:1279-1281)

There are numerous approaches to inhibiting PCSK9 being developed as a means of lowering cholesterol levels (reviewed in Lambert et al., 2012; J Lipid Res. 53(12):2515-24). A number of monoclonal antibodies that bind to PCSK9 near the catalytic domain that interact with the LDLR and hence inhibit the function of PCSK9 are currently in clinical trials including AMG145, 1D05-IgG2, and SAR236553/REGN727 (Aventis/Regeneron). Peptide mimetics and oligonucleotide approaches are also being developed. These include a mimic of the EGFA domain of the LDLR that binds to PCSK9, an antisense PCSK9 oligonucleotide, a locked nucleic acid inhibitor and siRNA approaches.

Protein: PCSK9 Gene: PCSK9 (Homo sapiens, chromosome 1, 55505149-55530526 [NCBI Reference Sequence: NC_(—)000001.10]; start site location: 55505511; strand: positive)

Gene Identification GeneID 255738 HGNC 20001 HPRD 07080 MIM 607789

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 5235 CAGGGCGCGTGAAGGGGCGCGCGG 120 5236 GACGCGTCCCGGCCCGCCCGAGC 179 5285 GACGCCTGGGGCGCGCAGATCAC 341 5341 CAGGCCGGCGCCCTAGGGGCTCC 494 5359 CACGCCGGCGGCGCCTTGAGCC 56 5402 PC2 CAGGTTTCGGCCTCGCCCTCCC 408 5445 CATCGAGCCCGCCATCGCAGCAC 1307 5473 GAGCGCCTCGACGTCGCTGCGGAAACC 273

Target Shift Sequence Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 5235 CAGGGCGCGTGAAGGGGCGCGCGG 120 5236 GACGCGTCCCGGCCCGCCCGAGC 179 5237 ACGCGTCCCGGCCCGCCCGA 180 5238 CGCGTCCCGGCCCGCCCGAG 181 5239 GCGTCCCGGCCCGCCCGAGC 182 5240 CGTCCCGGCCCGCCCGAGCC 183 5241 GTCCCGGCCCGCCCGAGCCA 184 5242 TCCCGGCCCGCCCGAGCCAG 185 5243 CCCGGCCCGCCCGAGCCAGT 186 5244 CCGGCCCGCCCGAGCCAGTC 187 5245 CGGCCCGCCCGAGCCAGTCT 188 5246 GGCCCGCCCGAGCCAGTCTC 189 5247 GCCCGCCCGAGCCAGTCTCA 190 5248 CCCGCCCGAGCCAGTCTCAC 191 5249 CCGCCCGAGCCAGTCTCACT 192 5250 CGCCCGAGCCAGTCTCACTG 193 5251 GCCCGAGCCAGTCTCACTGC 194 5252 CCCGAGCCAGTCTCACTGCC 195 5253 CCGAGCCAGTCTCACTGCCT 196 5254 CGAGCCAGTCTCACTGCCTG 197 5255 CGACGCGTCCCGGCCCGCCC 178 5256 ACGACGCGTCCCGGCCCGCC 177 5257 AACGACGCGTCCCGGCCCGC 176 5258 CAACGACGCGTCCCGGCCCG 175 5259 GCAACGACGCGTCCCGGCCC 174 5260 TGCAACGACGCGTCCCGGCC 173 5261 CTGCAACGACGCGTCCCGGC 172 5262 GCTGCAACGACGCGTCCCGG 171 5263 TGCTGCAACGACGCGTCCCG 170 5264 CTGCTGCAACGACGCGTCCC 169 5265 GCTGCTGCAACGACGCGTCC 168 5266 CGCTGCTGCAACGACGCGTC 167 5267 CCGCTGCTGCAACGACGCGT 166 5268 GCCGCTGCTGCAACGACGCG 165 5269 AGCCGCTGCTGCAACGACGC 164 5270 GAGCCGCTGCTGCAACGACG 163 5271 GGAGCCGCTGCTGCAACGAC 162 5272 GGGAGCCGCTGCTGCAACGA 161 5273 TGGGAGCCGCTGCTGCAACG 160 5274 CTGGGAGCCGCTGCTGCAAC 159 5275 GCTGGGAGCCGCTGCTGCAA 158 5276 AGCTGGGAGCCGCTGCTGCA 157 5277 GAGCTGGGAGCCGCTGCTGC 156 5278 GGAGCTGGGAGCCGCTGCTG 155 5279 GGGAGCTGGGAGCCGCTGCT 154 5280 TGGGAGCTGGGAGCCGCTGC 153 5281 CTGGGAGCTGGGAGCCGCTG 152 5282 GCTGGGAGCTGGGAGCCGCT 151 5283 GGCTGGGAGCTGGGAGCCGC 150 5284 TGGCTGGGAGCTGGGAGCCG 149 5285 GACGCCTGGGGCGCGCAGATCAC 319 5286 ACGCCTGGGGCGCGCAGATC 320 5287 CGCCTGGGGCGCGCAGATCA 321 5288 GCCTGGGGCGCGCAGATCAC 322 5289 CCTGGGGCGCGCAGATCACG 323 5290 CTGGGGCGCGCAGATCACGC 324 5291 TGGGGCGCGCAGATCACGCC 325 5292 GGGGCGCGCAGATCACGCCA 326 5293 GGGCGCGCAGATCACGCCAC 327 5294 GGCGCGCAGATCACGCCACC 328 5295 GCGCGCAGATCACGCCACCA 329 5296 CGCGCAGATCACGCCACCAG 330 5297 GCGCAGATCACGCCACCAGA 331 5298 CGCAGATCACGCCACCAGAG 332 5299 GCAGATCACGCCACCAGAGC 333 5300 CAGATCACGCCACCAGAGCC 334 5301 AGATCACGCCACCAGAGCCC 335 5302 GATCACGCCACCAGAGCCCC 336 5303 ATCACGCCACCAGAGCCCCA 337 5304 TCACGCCACCAGAGCCCCAT 338 5305 CACGCCACCAGAGCCCCATC 339 5306 ACGCCACCAGAGCCCCATCG 340 5307 CGCCACCAGAGCCCCATCGG 341 5308 GCCACCAGAGCCCCATCGGA 342 5309 CCACCAGAGCCCCATCGGAC 343 5310 CACCAGAGCCCCATCGGACG 344 5311 ACCAGAGCCCCATCGGACGA 345 5312 CCAGAGCCCCATCGGACGAT 346 5313 CAGAGCCCCATCGGACGATC 347 5314 AGAGCCCCATCGGACGATCC 348 5315 GAGCCCCATCGGACGATCCT 349 5316 AGCCCCATCGGACGATCCTA 350 5317 GCCCCATCGGACGATCCTAT 351 5318 CCCCATCGGACGATCCTATC 352 5319 CCCATCGGACGATCCTATCT 353 5320 CCATCGGACGATCCTATCTG 354 5321 CATCGGACGATCCTATCTGA 355 5322 ATCGGACGATCCTATCTGAT 356 5323 TCGGACGATCCTATCTGATT 357 5324 CGGACGATCCTATCTGATTA 358 5325 TGACGCCTGGGGCGCGCAGA 318 5326 TTGACGCCTGGGGCGCGCAG 317 5327 CTTGACGCCTGGGGCGCGCA 316 5328 GCTTGACGCCTGGGGCGCGC 315 5329 TGCTTGACGCCTGGGGCGCG 314 5330 GTGCTTGACGCCTGGGGCGC 313 5331 GGTGCTTGACGCCTGGGGCG 312 5332 GGGTGCTTGACGCCTGGGGC 311 5333 TGGGTGCTTGACGCCTGGGG 310 5334 GTGGGTGCTTGACGCCTGGG 309 5335 TGTGGGTGCTTGACGCCTGG 308 5336 GTGTGGGTGCTTGACGCCTG 307 5337 GGTGTGGGTGCTTGACGCCT 306 5338 GGGTGTGGGTGCTTGACGCC 305 5339 AGGGTGTGGGTGCTTGACGC 304 5340 TAGGGTGTGGGTGCTTGACG 303 5341 CAGGCCGGCGCCCTAGGGGCTCC 494 5342 AGGCCGGCGCCCTAGGGGCT 495 5343 GGCCGGCGCCCTAGGGGCTC 496 5344 GCCGGCGCCCTAGGGGCTCC 497 5345 CCGGCGCCCTAGGGGCTCCT 498 5346 CGGCGCCCTAGGGGCTCCTC 499 5347 GGCGCCCTAGGGGCTCCTCC 500 5348 GCGCCCTAGGGGCTCCTCCT 501 5349 CGCCCTAGGGGCTCCTCCTC 502 5350 GCAGGCCGGCGCCCTAGGGG 493 5351 GGCAGGCCGGCGCCCTAGGG 492 5352 AGGCAGGCCGGCGCCCTAGG 491 5353 AAGGCAGGCCGGCGCCCTAG 490 5354 GAAGGCAGGCCGGCGCCCTA 489 5355 GGAAGGCAGGCCGGCGCCCT 488 5356 TGGAAGGCAGGCCGGCGCCC 487 5357 CTGGAAGGCAGGCCGGCGCC 486 5358 GCTGGAAGGCAGGCCGGCGC 485 5359 CACGCCGGCGGCGCCTTGAGCC 58 5360 ACGCCGGCGGCGCCTTGAGC 59 5361 CGCCGGCGGCGCCTTGAGCC 60 5362 GCCGGCGGCGCCTTGAGCCT 61 5363 CCGGCGGCGCCTTGAGCCTT 62 5364 CGGCGGCGCCTTGAGCCTTG 63 5365 GGCGGCGCCTTGAGCCTTGC 64 5366 GCGGCGCCTTGAGCCTTGCG 65 5367 CGGCGCCTTGAGCCTTGCGG 66 5368 GGCGCCTTGAGCCTTGCGGT 67 5369 GCGCCTTGAGCCTTGCGGTG 68 5370 CGCCTTGAGCCTTGCGGTGG 69 5371 GCCTTGAGCCTTGCGGTGGG 70 5372 CCTTGAGCCTTGCGGTGGGG 71 5373 CTTGAGCCTTGCGGTGGGGA 72 5374 TTGAGCCTTGCGGTGGGGAG 73 5375 TGAGCCTTGCGGTGGGGAGG 74 5376 CCACGCCGGCGGCGCCTTGA 57 5377 TCCACGCCGGCGGCGCCTTG 56 5378 GTCCACGCCGGCGGCGCCTT 55 5379 GGTCCACGCCGGCGGCGCCT 54 5380 CGGTCCACGCCGGCGGCGCC 53 5381 GCGGTCCACGCCGGCGGCGC 52 5382 CGCGGTCCACGCCGGCGGCG 51 5383 GCGCGGTCCACGCCGGCGGC 50 5384 TGCGCGGTCCACGCCGGCGG 49 5385 GTGCGCGGTCCACGCCGGCG 48 5386 CGTGCGCGGTCCACGCCGGC 47 5387 CCGTGCGCGGTCCACGCCGG 46 5388 GCCGTGCGCGGTCCACGCCG 45 5389 GGCCGTGCGCGGTCCACGCC 44 5390 AGGCCGTGCGCGGTCCACGC 43 5391 GAGGCCGTGCGCGGTCCACG 42 5392 AGAGGCCGTGCGCGGTCCAC 41 5393 TAGAGGCCGTGCGCGGTCCA 40 5394 CTAGAGGCCGTGCGCGGTCC 39 5395 CCTAGAGGCCGTGCGCGGTC 38 5396 ACCTAGAGGCCGTGCGCGGT 37 5397 GACCTAGAGGCCGTGCGCGG 36 5398 AGACCTAGAGGCCGTGCGCG 35 5399 GAGACCTAGAGGCCGTGCGC 34 5400 GGAGACCTAGAGGCCGTGCG 33 5401 AGGAGACCTAGAGGCCGTGC 32 5402 CAGGTTTCGGCCTCGCCCTCCC 408 5403 AGGTTTCGGCCTCGCCCTCC 409 5404 GGTTTCGGCCTCGCCCTCCC 410 5405 GTTTCGGCCTCGCCCTCCCC 411 5406 TTTCGGCCTCGCCCTCCCCA 412 5407 TTCGGCCTCGCCCTCCCCAA 413 5408 TCGGCCTCGCCCTCCCCAAA 414 5409 CGGCCTCGCCCTCCCCAAAC 415 5410 GGCCTCGCCCTCCCCAAACA 416 5411 GCCTCGCCCTCCCCAAACAG 417 5412 CCTCGCCCTCCCCAAACAGC 418 5413 CTCGCCCTCCCCAAACAGCG 419 5414 TCGCCCTCCCCAAACAGCGT 420 5415 CGCCCTCCCCAAACAGCGTC 421 5416 GCCCTCCCCAAACAGCGTCA 422 5417 CCCTCCCCAAACAGCGTCAG 423 5418 CCTCCCCAAACAGCGTCAGA 424 5419 CTCCCCAAACAGCGTCAGAT 425 5420 TCCCCAAACAGCGTCAGATT 426 5421 CCCCAAACAGCGTCAGATTA 427 5422 CCCAAACAGCGTCAGATTAC 428 5423 CCAAACAGCGTCAGATTACG 429 5424 CAAACAGCGTCAGATTACGC 430 5425 AAACAGCGTCAGATTACGCG 431 5426 AACAGCGTCAGATTACGCGC 432 5427 ACAGCGTCAGATTACGCGCA 433 5428 CAGCGTCAGATTACGCGCAG 434 5429 AGCGTCAGATTACGCGCAGA 435 5430 GCGTCAGATTACGCGCAGAG 436 5431 CGTCAGATTACGCGCAGAGG 437 5432 GTCAGATTACGCGCAGAGGG 438 5433 TCAGATTACGCGCAGAGGGA 439 5434 TCAGGTTTCGGCCTCGCCCT 407 5435 ATCAGGTTTCGGCCTCGCCC 406 5436 GATCAGGTTTCGGCCTCGCC 405 5437 GGATCAGGTTTCGGCCTCGC 404 5438 AGGATCAGGTTTCGGCCTCG 403 5439 GAGGATCAGGTTTCGGCCTC 402 5440 GGAGGATCAGGTTTCGGCCT 401 5441 TGGAGGATCAGGTTTCGGCC 400 5442 CTGGAGGATCAGGTTTCGGC 399 5443 ACTGGAGGATCAGGTTTCGG 398 5444 GACTGGAGGATCAGGTTTCG 397 5445 CATCGAGCCCGCCATCGCAGCAC 1307 5446 ATCGAGCCCGCCATCGCAGC 1308 5447 TCGAGCCCGCCATCGCAGCA 1309 5448 CGAGCCCGCCATCGCAGCAC 1310 5449 GAGCCCGCCATCGCAGCACA 1311 5450 AGCCCGCCATCGCAGCACAG 1312 5451 GCCCGCCATCGCAGCACAGA 1313 5452 CCCGCCATCGCAGCACAGAG 1314 5453 CCGCCATCGCAGCACAGAGT 1315 5454 CGCCATCGCAGCACAGAGTA 1316 5455 GCCATCGCAGCACAGAGTAG 1317 5456 CCATCGCAGCACAGAGTAGG 1318 5457 CATCGCAGCACAGAGTAGGA 1319 5458 CCATCGAGCCCGCCATCGCA 1306 5459 CCCATCGAGCCCGCCATCGC 1305 5460 CCCCATCGAGCCCGCCATCG 1304 5461 TCCCCATCGAGCCCGCCATC 1303 5462 ATCCCCATCGAGCCCGCCAT 1302 5463 TATCCCCATCGAGCCCGCCA 1301 5464 TTATCCCCATCGAGCCCGCC 1300 5465 GTTATCCCCATCGAGCCCGC 1299 5466 AGTTATCCCCATCGAGCCCG 1298 5467 GAGTTATCCCCATCGAGCCC 1297 5468 AGAGTTATCCCCATCGAGCC 1296 5469 CAGAGTTATCCCCATCGAGC 1295 5470 TCAGAGTTATCCCCATCGAG 1294 5471 GTCAGAGTTATCCCCATCGA 1293 5472 GGTCAGAGTTATCCCCATCG 1292 5473 GAGCGCCTCGACGTCGCTGCGGAAACC 273 5474 AGCGCCTCGACGTCGCTGCG 274 5475 GCGCCTCGACGTCGCTGCGG 275 5476 CGCCTCGACGTCGCTGCGGA 276 5477 GCCTCGACGTCGCTGCGGAA 277 5478 CCTCGACGTCGCTGCGGAAA 278 5479 CTCGACGTCGCTGCGGAAAC 279 5480 TCGACGTCGCTGCGGAAACC 280 5481 CGACGTCGCTGCGGAAACCT 281 5482 GACGTCGCTGCGGAAACCTT 282 5483 ACGTCGCTGCGGAAACCTTC 283 5484 CGTCGCTGCGGAAACCTTCT 284 5485 GTCGCTGCGGAAACCTTCTA 285 5486 TCGCTGCGGAAACCTTCTAG 286 5487 CGCTGCGGAAACCTTCTAGG 287 5488 GCTGCGGAAACCTTCTAGGG 288 5489 CTGCGGAAACCTTCTAGGGT 289 5490 TGCGGAAACCTTCTAGGGTG 290 5491 GCGGAAACCTTCTAGGGTGT 291 5492 CGGAAACCTTCTAGGGTGTG 292 5493 TGAGCGCCTCGACGTCGCTG 272 5494 ATGAGCGCCTCGACGTCGCT 271 5495 CATGAGCGCCTCGACGTCGC 270 5496 CCATGAGCGCCTCGACGTCG 269 5497 ACCATGAGCGCCTCGACGTC 268 5498 AACCATGAGCGCCTCGACGT 267 5499 CAACCATGAGCGCCTCGACG 266 5500 GCAACCATGAGCGCCTCGAC 265 5501 TGCAACCATGAGCGCCTCGA 264 5502 CTGCAACCATGAGCGCCTCG 263 5503 CCTGCAACCATGAGCGCCTC 262 5504 GCCTGCAACCATGAGCGCCT 261 5505 CGCCTGCAACCATGAGCGCC 260 5506 CCGCCTGCAACCATGAGCGC 259 5507 CCCGCCTGCAACCATGAGCG 258 5508 GCCCGCCTGCAACCATGAGC 257 5509 CGCCCGCCTGCAACCATGAG 256 5510 GCGCCCGCCTGCAACCATGA 255 5511 GGCGCCCGCCTGCAACCATG 254 5512 CGGCGCCCGCCTGCAACCAT 253 5513 GCGGCGCCCGCCTGCAACCA 252 5514 GGCGGCGCCCGCCTGCAACC 251 5515 CGGCGGCGCCCGCCTGCAAC 250 5516 ACGGCGGCGCCCGCCTGCAA 249 5517 AACGGCGGCGCCCGCCTGCA 248 5518 GAACGGCGGCGCCCGCCTGC 247 5519 TGAACGGCGGCGCCCGCCTG 246 5520 CTGAACGGCGGCGCCCGCCT 245 5521 ACTGAACGGCGGCGCCCGCC 244 5522 AACTGAACGGCGGCGCCCGC 243 5523 GAACTGAACGGCGGCGCCCG 242 5524 TGAACTGAACGGCGGCGCCC 241 5525 CTGAACTGAACGGCGGCGCC 240 5526 CCTGAACTGAACGGCGGCGC 239 5527 CCCTGAACTGAACGGCGGCG 238 5528 ACCCTGAACTGAACGGCGGC 237 5529 GACCCTGAACTGAACGGCGG 236 5530 AGACCCTGAACTGAACGGCG 235 5531 CAGACCCTGAACTGAACGGC 234 5532 TCAGACCCTGAACTGAACGG 233 5533 CTCAGACCCTGAACTGAACG 232

Hot Zones (Relative upstream location to gene start site)  1-800 1100-1450

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11968) CCATCCTGGGCCATTGGGCCAGCTCCAGCCTCATCCTTGAATGGTGGGTG TACATCGCTGGGGTCCTTGCTAGATTATATTAGGGCCTCCACACACTTTA GTTGCTCTCCGTTTGCACAATGATTTCTATATCATTAGTACCTCTTTGTC CACTTGCCTAATTATTTCCTTAGGATAAATTCCTAAGAAATCAAATAGCT AGGTCAGTATATAGCACATTTTCACAGTTTGCTACTGACTAGAGATATTA AAGGTACAAAAACAGCCAGAATTAGATGTAGAGTCGCAAGGGAGTTTTGT TACTAAGACGTCATTTCTAATCAGGGAGAAAAAATGAGTTACGGAATAAA CGGTATTGGAACAATAGGCTAGCCTTCTGGAGAAAAATAACCTCACTCCT TATACAAAAGTAAATCCCAGTGGAAGCCAAGATATTAAAAAAAGATTTAA AAAAGGAGGACAATTTTTATATGATCTTGATAAGGAGGAGGCCTTTCTAA GCACAGTACAAAATCAAGAAGTCATGAAATAAAAGGCTGATAAGTTTGAC TCCATGAAAATTAAAATTTTCTATGGAAAAAAATACAATAAAGTCAAAAA TCAATCAACAAACTGGAAAATAGATTTGCAACATACATAAGAGACAGCAG GCTAATTTTGGTATTATACATAAAAAGCTATTACAAATCATCAAACAAAA GCTCCACAGCCAAAATGAAAAAATGAAGAGACAGTTCAAAGACAAACAAA TGTAGATAATTGTTTAACTTAAGGAAAGGTCAAAATATTTGGCAACTCTG TGTTGGCCTGGGTGTGACCATGAGCGGTAGTTGCCAGTGGTATTCACAAA TATACCCTTTCTCCTCCTTCTGGGCACTTGGTGTGATTGCAGTTTCCTAC ATTTGACCTTAGGTGTGGTCATGTGTCTCGCTTAGGAGAAGGAAATGTGA ACGGAAGTGTTGTGGGTCACTTTTGTGTGGAAGCTGCATGGAGTCACCTC TTTGCTTTCCGTCAGCCTCAGTGTTCAGCAATGTTCTGAATGATGGTTGC TTTATCAGTCTGGTTCTGGGGTGAGGGTAATGAAGTAGTGAAGCAAAGTC CTTGATGGACATGTGAGGTGAGACAGAAATAAACCTTTGACATTTCAAGC CCCTGAGATTTGGGGCGTGCATGTGCTGGAAGCAGAACCTAGCCTATTCT GATGGATTCCTCCAGCACTGCTCGTGGGAAGACATCATCAATATAGAGCA TTCATGTGCCCTTTATCTCGAAATTCCACTTCCAGGAATTTATGAAACAG ATACTCTCACATGTGCAAACAGCTATGGATAAGGACAGACATGGCAACTT GGATTGCAATAGCAAAAGACAGAAACAACCAACGGAAACACCAACCAATA GGAAATTGGCTAAAGACATTGTGAAACATACATAGAATGAAATAATCTGC AACCAGAAAAATAAGGCAGTAGATGTATGTGTACCAGTGTGGTTTTTATT CCGAGATTAGGGCTAGGTTAAGACGTCAGATTAAGTTGTCCCTCTCCACC CCACCAATATAAATAAAAAGTTAAAAGTAAATCATAAACTATTTTTACAA TTTTAAAAAGTGGGTTAAAGAGCCCATCCAAGTAGTTTTATAAAAGTAGA CTATCTCCGAAAAGATACCCAATAAATAGGTATATTACTTTCCTGGGGCT GTTATAACAAGTTTCTACAAATTTGCTGGCTTCAAATAACAAAAACGTAT TCTCTTGCAGTTATGGAAGCCAGAAGTATGGAATGAAGGGTTGCAGGGTG GTGCCCTCTCCCAAAGCTCTAGGGGAGGAACATTCCTTGCTTCTTCCAGC TCCTTTGGGGGCTCCTGGCATTCCTTGGCTTATGTCGGCACAGCTCTAAT CGGCGCCTCCATTGTTACATAGGTGTTTCTGTGTCTCAAGTATCTCTCCC CTTTCTCTTCTGATATCAGTCATTGGATTTAGGGACCATCCTAAACCCAG GATAATCTCCTCATGAGATCCTTAGGTCAATTACATCTGCAAAGATCTCA TTTCCAAATAAGGTCACATTCAAAAGTACCAGGGGTTAGTCTTAGACTTA TCTTTTTGGGGGACACGATTCAACCCACTACCGTGGGTAACAGTGGTTTT CCCTCAGAAGGTGGTGGTTCAGGAGTGGGAGGAAGATGAACTTTTCACTG TATATCCTTTCAAACTATTCACGTTTTAAAAAAAACATTTTCATGTAAAT TTAAAAAAATTGAACATTCACACAAAAAGATGCCCCCTCCCTTGCAAAAA AGAGTATGCCCGTTCAAAATGTTGAAATGTACACTCACAGCAATGGTGGC TGCAGACTCCAAGTTTCTGAGGTTGGAGAAGGTAGCCAGGGAGCATAAAA GTGAGTTCTATCTACTCATTCAGTCTATGAGGGGAAGGCAATGGCTAGAA AAGCATTTTGAGGGACAGTAAAAGTGGCATTTTTAGAGGGAGGAAGCCTT GAGGATGCTTGTGGGGTGAAGGGAAAGAATAACTCAGGAAGAGGCATTTA GGGATAAGAGGAGGAGAGGAGATAGTGGAGGTAGGTGATCCCTGCGGAGG CCAGATTGGGGCAGGGGAGTGTCAGCTGAGTATAAGAGGATGGTCCCCTC TGCCCTGAAGGAGGAAGGCAGGAGGGGAAAAGGATGGGTGTTGACCCAGA AAGCACTTGTGGTGGAGGGGAGGCCCCAGAAGAGGCTTCTGACTTACCCT GATTGCTGGTACCTCTCAGGGGAGCTGGCTGCTTATTTGCTGGCCAGGGT GTGGGGGAACCCATTTGAGAAGAGGGAGAAGGTGACACAATTCCTTTGGG CAACTTATGGGAGGGGTAATTGGTGAGGGATGAAAGCCCTGCCAAGTGGC AGGAGGCCCAGCTGGGGCTGCCCCTCATAAGAGTGCAGTGGAGGATATGG GATGAGAAGTGACTGCCCCTCTGGTTCCATCTGTCGCAGAGCCCAGGGTG CTTCCTTCCTCCCCCACCTCCCTCAGAACACACCCACTGCATGCTGGACA GCAGCCCCCTTCCTGGGCCTGGGGACATCCATGTCCCTCTGTGCACAGGC TTCATCATTCTCTGGGTGCACGGTAACGACCCCGGTAGGTGAGAGGCCAA GGTCCCAAAGGGGAGCAGCAGGGAAAGTTAGCTCCCATCTATTCTTGCTC CAGGGGAGGCCTTTGATGAGGAAGCTGCCAAAAGCACATTGCAAATACAA TTCCAATTACAGGCAACAGGAAGGAGAACCACCTCTGCCACCTCTGTCAG CAAACCATGAGCTCCTACTCTGTGCTGCGATGGCGGGCTCGATGGGGATA ACTCTGACCTTACCTCATGGAGTCACTGTCAACCCACTGGTTGCACTGTC TTTGTGCACTGGCTCTCTGGAGTGAGGTCTTTGCAAACAAAGTGGAAAGA GCATCAACTTTGGACTCCAGCACCTAGATTCAGAGCAGGCCATTTCACTC GGAATCTGCTGTGCATCTGCAAGGGAGGATCATAAATTCGCCTTTGTTTC TTCCCAGTATCGACAGCCCTTCCAGAAAGAGCAAGCCTCATGTCATGCCA CATGTACAATCTGAGGCCAGGAGCTCTCTTTCCCCTTTTCATCCTCCTGC CTGGTACACAATAGGTGTTTACTGGATGCTTGTCCAGTTGATTTCTTGAA CATGGTGTGTAAAAGGAATCTTTGCAAATTGAATCTTCTGGAAAGCTGAG CTTGTGCCTACCATAGAATTCTGAATGTACCTATATGACGTCTTTGCAAA CTTAAAACCTGAATCTTTGTAGTATAAATCCCTTGAAATGCATGTAGGCT GGACATCAAAAGCAAGCAATCTCTTCAAGGAGCAGCTAGTTGGTAAGGTC AGTGTGCAGGGTGCATAAAGGGCAGAGGCCGGAGGGGGTCCAGGCTAAGT TTAGAAGGCTGCCAGGTTAAGGCCAGTGGAAAGAATTCGGTGGGCAGCGA GGAGTCCACAGTAGGATTGATTCAGAAGTCTCACTGGTCAGCAGGAGACA AGGTGGACCCAGGAAACACTGAAAAGGTGGGCCCGGCAGAACTTGGAGTC TGGCATCCCACGCAGGGTGAGAGGCGGGAGAGGAGGAGCCCCTAGGGCGC CGGCCTGCCTTCCAGCCCAGTTAGGATTTGGGAGTTTTTTCTTCCCTCTG CGCGTAATCTGACGCTGTTTGGGGAGGGCGAGGCCGAAACCTGATCCTCC AGTCCGGGGGTTCCGTTAATGTTTAATCAGATAGGATCGTCCGATGGGGC TCTGGTGGCGTGATCTGCGCGCCCCAGGCGTCAAGCACCCACACCCTAGA AGGTTTCCGCAGCGACGTCGAGGCGCTCATGGTTGCAGGCGGGCGCCGCC GTTCAGTTCAGGGTCTGAGCCTGGAGGAGTGAGCCAGGCAGTGAGACTGG CTCGGGCGGGCCGGGACGCGTCGTTGCAGCAGCGGCTCCCAGCTCCCAGC CAGGATTCCGCGCGCCCCTTCACGCGCCCTGCTCCTGAACTTCAGCTCCT GCACAGTCCTCCCCACCGCAAGGCTCAAGGCGCCGCCGGCGTGGACCGCG CACGGCCTCTAGGTCTCCTCGCCAGGACAGCAACCTCTCCCCTGGCCCTC ATG

20) MEK1. MEK1 (MAP2K1) Mitogen-activated protein kinase kinase 1. Dual specificity protein kinases act as an essential component of the MAP kinase signal transduction pathway and serves as an integration point for multiple biochemical signals. MEK1 and MEK2 are members of the dual specificity protein kinase family, which act as a mitogen-activated protein (MAP) kinase kinases and as extracellular signal-regulated kinases (ERKs). Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. RAF1 then further activates the dual-specificity protein kinases MAP2K1/MEK1 and MAP2K2/MEK2. Both MAP2K1/MEK1 and MAP2K2/MEK2 function specifically in the MAPK/ERK cascade, and catalyze the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, leading to their activation and further transduction of the signal within the MAPK/ERK cascade. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth and proliferation, adhesion, survival and differentiation, predominantly through the regulation of transcription, metabolism and cytoskeletal rearrangements (reviewed by Roberts and Der; 2007 Oncogene 26, 3291-3310).

Genetic alterations that activate the mitogen-activated protein kinase (MAP kinase) pathway occur commonly in cancer. For example, the majority of melanomas harbor mutations in the BRAF oncogene, which confers enhanced sensitivity to pharmacologic MAP kinase inhibition (e.g., RAF or MEK inhibitors). Most mutations conferring resistance to MEK inhibition in vitro populated the allosteric drug binding pocket or alpha-helix C and showed robust (approximately 100-fold) resistance to allosteric MEK inhibition (reviewed in Emery et al, 2009; Proc Natl Acad Sci.; 106(48):20411-20416). Other mutations affected MEK1 codons located within or abutting the N-terminal negative regulatory helix (helix A), which also undergo gain-of-function germline mutations in cardiofaciocutaneous (CFC) syndrome. One target of the MAPK/ERK cascade is peroxisome proliferator-activated receptor gamma (PPARG), a nuclear receptor that promotes differentiation and apoptosis. MAP2K1/MEK1 has been shown to export PPARG from the nucleus. The MAPK/ERK cascade is also involved in the regulation of endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC), as well as in the fragmentation of the Golgi apparatus during mitosis.

Protein: MEK1 Gene: MAP2K1 (Homo sapiens, chromosome 15, 66679211-66783882 [NCBI Reference Sequence: NC_(—)000015.9]; start site location: 66679686; strand: positive)

Gene Identification GeneID 5604 HGNC 6840 HPRD 01469 MIM 176872

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 5534 CAAGTCCGGGCCGCGGGCCCCGGGGC 93 5716 MEK1_2 GCGCCCCGCGCGGTCCCGTCAGCGC 133 5898 GCGGAGCGGGCTGAACGTGCG 249 5900 GACTGGAGGCCGGGGGAGGGGCGGGG 433 5901 GACCCGGGTAACGCGCTTCCAAC 5 5924 MEK1_1 CACTCGGCTCCGCCCCTATTGC 507 6000 TACGTCACGGGAGCGCGGCGCAC 578 6077 GTCGCGGACGCCGTGGCGCCCTCTGTC 619 6154 CACTCGCCGTCATGCCCGGATCC 1183

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 5534 CAAGTCCGGGCCGCGGGCCCCGGGGC 93 5535 AAGTCCGGGCCGCGGGCCCC 94 5536 AGTCCGGGCCGCGGGCCCCG 95 5537 GTCCGGGCCGCGGGCCCCGG 96 5538 TCCGGGCCGCGGGCCCCGGG 97 5539 CCGGGCCGCGGGCCCCGGGG 98 5540 CGGGCCGCGGGCCCCGGGGC 99 5541 GGGCCGCGGGCCCCGGGGCT 100 5542 GGCCGCGGGCCCCGGGGCTG 101 5543 GCCGCGGGCCCCGGGGCTGC 102 5544 CCGCGGGCCCCGGGGCTGCC 103 5545 CGCGGGCCCCGGGGCTGCCT 104 5546 GCGGGCCCCGGGGCTGCCTT 105 5547 CGGGCCCCGGGGCTGCCTTC 106 5548 GGGCCCCGGGGCTGCCTTCA 107 5549 GGCCCCGGGGCTGCCTTCAG 108 5550 GCCCCGGGGCTGCCTTCAGC 109 5551 CCCCGGGGCTGCCTTCAGCG 110 5552 CCCGGGGCTGCCTTCAGCGG 111 5553 CCGGGGCTGCCTTCAGCGGG 112 5554 CGGGGCTGCCTTCAGCGGGT 113 5555 GGGGCTGCCTTCAGCGGGTG 114 5556 GGGCTGCCTTCAGCGGGTGC 115 5557 GGCTGCCTTCAGCGGGTGCG 116 5558 GCTGCCTTCAGCGGGTGCGC 117 5559 CTGCCTTCAGCGGGTGCGCC 118 5560 TGCCTTCAGCGGGTGCGCCC 119 5561 GCCTTCAGCGGGTGCGCCCC 120 5562 CCTTCAGCGGGTGCGCCCCG 121 5563 CTTCAGCGGGTGCGCCCCGC 122 5564 TTCAGCGGGTGCGCCCCGCG 123 5565 TCAGCGGGTGCGCCCCGCGC 124 5566 CAGCGGGTGCGCCCCGCGCG 125 5567 AGCGGGTGCGCCCCGCGCGG 126 5568 GCGGGTGCGCCCCGCGCGGT 127 5569 CGGGTGCGCCCCGCGCGGTC 128 5570 GGGTGCGCCCCGCGCGGTCC 129 5571 GGTGCGCCCCGCGCGGTCCC 130 5572 GTGCGCCCCGCGCGGTCCCG 131 5573 TGCGCCCCGCGCGGTCCCGT 132 5574 GCGCCCCGCGCGGTCCCGTC 133 5575 CGCCCCGCGCGGTCCCGTCA 134 5576 GCCCCGCGCGGTCCCGTCAG 135 5577 CCCCGCGCGGTCCCGTCAGC 136 5578 CCCGCGCGGTCCCGTCAGCG 137 5579 CCGCGCGGTCCCGTCAGCGC 138 5580 CGCGCGGTCCCGTCAGCGCC 139 5581 GCGCGGTCCCGTCAGCGCCG 140 5582 CGCGGTCCCGTCAGCGCCGA 141 5583 GCGGTCCCGTCAGCGCCGAG 142 5584 CGGTCCCGTCAGCGCCGAGG 143 5585 GGTCCCGTCAGCGCCGAGGG 144 5586 GTCCCGTCAGCGCCGAGGGG 145 5587 TCCCGTCAGCGCCGAGGGGC 146 5588 CCCGTCAGCGCCGAGGGGCC 147 5589 CCGTCAGCGCCGAGGGGCCG 148 5590 CGTCAGCGCCGAGGGGCCGG 149 5591 GTCAGCGCCGAGGGGCCGGT 150 5592 TCAGCGCCGAGGGGCCGGTA 151 5593 CAGCGCCGAGGGGCCGGTAG 152 5594 AGCGCCGAGGGGCCGGTAGC 153 5595 GCGCCGAGGGGCCGGTAGCG 154 5596 CGCCGAGGGGCCGGTAGCGG 155 5597 GCCGAGGGGCCGGTAGCGGT 156 5598 CCGAGGGGCCGGTAGCGGTC 157 5599 CGAGGGGCCGGTAGCGGTCT 158 5600 GAGGGGCCGGTAGCGGTCTC 159 5601 AGGGGCCGGTAGCGGTCTCA 160 5602 GGGGCCGGTAGCGGTCTCAG 161 5603 GGGCCGGTAGCGGTCTCAGT 162 5604 GGCCGGTAGCGGTCTCAGTG 163 5605 GCCGGTAGCGGTCTCAGTGG 164 5606 CCGGTAGCGGTCTCAGTGGA 165 5607 CGGTAGCGGTCTCAGTGGAC 166 5608 GGTAGCGGTCTCAGTGGACC 167 5609 GTAGCGGTCTCAGTGGACCC 168 5610 TAGCGGTCTCAGTGGACCCC 169 5611 AGCGGTCTCAGTGGACCCCC 170 5612 GCGGTCTCAGTGGACCCCCG 171 5613 CGGTCTCAGTGGACCCCCGC 172 5614 GGTCTCAGTGGACCCCCGCC 173 5615 GTCTCAGTGGACCCCCGCCC 174 5616 TCTCAGTGGACCCCCGCCCC 175 5617 CTCAGTGGACCCCCGCCCCA 176 5618 TCAGTGGACCCCCGCCCCAC 177 5619 CAGTGGACCCCCGCCCCACC 178 5620 AGTGGACCCCCGCCCCACCC 179 5621 GTGGACCCCCGCCCCACCCG 180 5622 TGGACCCCCGCCCCACCCGC 181 5623 GGACCCCCGCCCCACCCGCC 182 5624 GACCCCCGCCCCACCCGCCC 183 5625 ACCCCCGCCCCACCCGCCCG 184 5626 CCCCCGCCCCACCCGCCCGG 185 5627 CCCCGCCCCACCCGCCCGGG 186 5628 CCCGCCCCACCCGCCCGGGA 187 5629 CCGCCCCACCCGCCCGGGAC 188 5630 CGCCCCACCCGCCCGGGACT 189 5631 GCCCCACCCGCCCGGGACTC 190 5632 CCCCACCCGCCCGGGACTCG 191 5633 CCCACCCGCCCGGGACTCGG 192 5634 CCACCCGCCCGGGACTCGGC 193 5635 CACCCGCCCGGGACTCGGCT 194 5636 ACCCGCCCGGGACTCGGCTT 195 5637 CCCGCCCGGGACTCGGCTTC 196 5638 CCGCCCGGGACTCGGCTTCG 197 5639 CGCCCGGGACTCGGCTTCGC 198 5640 GCCCGGGACTCGGCTTCGCG 199 5641 CCCGGGACTCGGCTTCGCGC 200 5642 CCGGGACTCGGCTTCGCGCG 201 5643 CGGGACTCGGCTTCGCGCGC 202 5644 GGGACTCGGCTTCGCGCGCA 203 5645 GGACTCGGCTTCGCGCGCAG 204 5646 GACTCGGCTTCGCGCGCAGA 205 5647 ACTCGGCTTCGCGCGCAGAG 206 5648 CTCGGCTTCGCGCGCAGAGA 207 5649 TCGGCTTCGCGCGCAGAGAG 208 5650 CGGCTTCGCGCGCAGAGAGC 209 5651 GGCTTCGCGCGCAGAGAGCC 210 5652 GCTTCGCGCGCAGAGAGCCG 211 5653 CTTCGCGCGCAGAGAGCCGA 212 5654 TTCGCGCGCAGAGAGCCGAA 213 5655 TCGCGCGCAGAGAGCCGAAA 214 5656 CCAAGTCCGGGCCGCGGGCC 92 5657 ACCAAGTCCGGGCCGCGGGC 91 5658 GACCAAGTCCGGGCCGCGGG 90 5659 GGACCAAGTCCGGGCCGCGG 89 5660 AGGACCAAGTCCGGGCCGCG 88 5661 CAGGACCAAGTCCGGGCCGC 87 5662 GCAGGACCAAGTCCGGGCCG 86 5663 CGCAGGACCAAGTCCGGGCC 85 5664 GCGCAGGACCAAGTCCGGGC 84 5665 TGCGCAGGACCAAGTCCGGG 83 5666 CTGCGCAGGACCAAGTCCGG 82 5667 GCTGCGCAGGACCAAGTCCG 81 5668 CGCTGCGCAGGACCAAGTCC 80 5669 CCGCTGCGCAGGACCAAGTC 79 5670 CCCGCTGCGCAGGACCAAGT 78 5671 GCCCGCTGCGCAGGACCAAG 77 5672 CGCCCGCTGCGCAGGACCAA 76 5673 GCGCCCGCTGCGCAGGACCA 75 5674 CGCGCCCGCTGCGCAGGACC 74 5675 CCGCGCCCGCTGCGCAGGAC 73 5676 CCCGCGCCCGCTGCGCAGGA 72 5677 CCCCGCGCCCGCTGCGCAGG 71 5678 GCCCCGCGCCCGCTGCGCAG 70 5679 TGCCCCGCGCCCGCTGCGCA 69 5680 CTGCCCCGCGCCCGCTGCGC 68 5681 GCTGCCCCGCGCCCGCTGCG 67 5682 CGCTGCCCCGCGCCCGCTGC 66 5683 GCGCTGCCCCGCGCCCGCTG 65 5684 TGCGCTGCCCCGCGCCCGCT 64 5685 CTGCGCTGCCCCGCGCCCGC 63 5686 GCTGCGCTGCCCCGCGCCCG 62 5687 CGCTGCGCTGCCCCGCGCCC 61 5688 CCGCTGCGCTGCCCCGCGCC 60 5689 CCCGCTGCGCTGCCCCGCGC 59 5690 TCCCGCTGCGCTGCCCCGCG 58 5691 CTCCCGCTGCGCTGCCCCGC 57 5692 CCTCCCGCTGCGCTGCCCCG 56 5693 TCCTCCCGCTGCGCTGCCCC 55 5694 TTCCTCCCGCTGCGCTGCCC 54 5695 CTTCCTCCCGCTGCGCTGCC 53 5696 GCTTCCTCCCGCTGCGCTGC 52 5697 CGCTTCCTCCCGCTGCGCTG 51 5698 TCGCTTCCTCCCGCTGCGCT 50 5699 CTCGCTTCCTCCCGCTGCGC 49 5700 TCTCGCTTCCTCCCGCTGCG 48 5701 CTCTCGCTTCCTCCCGCTGC 47 5702 CCTCTCGCTTCCTCCCGCTG 46 5703 ACCTCTCGCTTCCTCCCGCT 45 5704 CACCTCTCGCTTCCTCCCGC 44 5705 GCACCTCTCGCTTCCTCCCG 43 5706 AGCACCTCTCGCTTCCTCCC 42 5707 CAGCACCTCTCGCTTCCTCC 41 5708 GCAGCACCTCTCGCTTCCTC 40 5709 GGCAGCACCTCTCGCTTCCT 39 5710 GGGCAGCACCTCTCGCTTCC 38 5711 AGGGCAGCACCTCTCGCTTC 37 5712 GAGGGCAGCACCTCTCGCTT 36 5713 GGAGGGCAGCACCTCTCGCT 35 5714 GGGAGGGCAGCACCTCTCGC 34 5715 GGGGAGGGCAGCACCTCTCG 33 5716 GCGCCCCGCGCGGTCCCGTCAGCGC 133 5717 CGCCCCGCGCGGTCCCGTCA 134 5718 GCCCCGCGCGGTCCCGTCAG 135 5719 CCCCGCGCGGTCCCGTCAGC 136 5720 CCCGCGCGGTCCCGTCAGCG 137 5721 CCGCGCGGTCCCGTCAGCGC 138 5722 CGCGCGGTCCCGTCAGCGCC 139 5723 GCGCGGTCCCGTCAGCGCCG 140 5724 CGCGGTCCCGTCAGCGCCGA 141 5725 GCGGTCCCGTCAGCGCCGAG 142 5726 CGGTCCCGTCAGCGCCGAGG 143 5727 GGTCCCGTCAGCGCCGAGGG 144 5728 GTCCCGTCAGCGCCGAGGGG 145 5729 TCCCGTCAGCGCCGAGGGGC 146 5730 CCCGTCAGCGCCGAGGGGCC 147 5731 CCGTCAGCGCCGAGGGGCCG 148 5732 CGTCAGCGCCGAGGGGCCGG 149 5733 GTCAGCGCCGAGGGGCCGGT 150 5734 TCAGCGCCGAGGGGCCGGTA 151 5735 CAGCGCCGAGGGGCCGGTAG 152 5736 AGCGCCGAGGGGCCGGTAGC 153 5737 GCGCCGAGGGGCCGGTAGCG 154 5738 CGCCGAGGGGCCGGTAGCGG 155 5739 GCCGAGGGGCCGGTAGCGGT 156 5740 CCGAGGGGCCGGTAGCGGTC 157 5741 CGAGGGGCCGGTAGCGGTCT 158 5742 GAGGGGCCGGTAGCGGTCTC 159 5743 AGGGGCCGGTAGCGGTCTCA 160 5744 GGGGCCGGTAGCGGTCTCAG 161 5745 GGGCCGGTAGCGGTCTCAGT 162 5746 GGCCGGTAGCGGTCTCAGTG 163 5747 GCCGGTAGCGGTCTCAGTGG 164 5748 CCGGTAGCGGTCTCAGTGGA 165 5749 CGGTAGCGGTCTCAGTGGAC 166 5750 GGTAGCGGTCTCAGTGGACC 167 5751 GTAGCGGTCTCAGTGGACCC 168 5752 TAGCGGTCTCAGTGGACCCC 169 5753 AGCGGTCTCAGTGGACCCCC 170 5754 GCGGTCTCAGTGGACCCCCG 171 5755 CGGTCTCAGTGGACCCCCGC 172 5756 GGTCTCAGTGGACCCCCGCC 173 5757 GTCTCAGTGGACCCCCGCCC 174 5758 TCTCAGTGGACCCCCGCCCC 175 5759 CTCAGTGGACCCCCGCCCCA 176 5760 TCAGTGGACCCCCGCCCCAC 177 5761 CAGTGGACCCCCGCCCCACC 178 5762 AGTGGACCCCCGCCCCACCC 179 5763 GTGGACCCCCGCCCCACCCG 180 5764 TGGACCCCCGCCCCACCCGC 181 5765 GGACCCCCGCCCCACCCGCC 182 5766 GACCCCCGCCCCACCCGCCC 183 5767 ACCCCCGCCCCACCCGCCCG 184 5768 CCCCCGCCCCACCCGCCCGG 185 5769 CCCCGCCCCACCCGCCCGGG 186 5770 CCCGCCCCACCCGCCCGGGA 187 5771 CCGCCCCACCCGCCCGGGAC 188 5772 CGCCCCACCCGCCCGGGACT 189 5773 GCCCCACCCGCCCGGGACTC 190 5774 CCCCACCCGCCCGGGACTCG 191 5775 CCCACCCGCCCGGGACTCGG 192 5776 CCACCCGCCCGGGACTCGGC 193 5777 CACCCGCCCGGGACTCGGCT 194 5778 ACCCGCCCGGGACTCGGCTT 195 5779 CCCGCCCGGGACTCGGCTTC 196 5780 CCGCCCGGGACTCGGCTTCG 197 5781 CGCCCGGGACTCGGCTTCGC 198 5782 GCCCGGGACTCGGCTTCGCG 199 5783 CCCGGGACTCGGCTTCGCGC 200 5784 CCGGGACTCGGCTTCGCGCG 201 5785 CGGGACTCGGCTTCGCGCGC 202 5786 GGGACTCGGCTTCGCGCGCA 203 5787 GGACTCGGCTTCGCGCGCAG 204 5788 GACTCGGCTTCGCGCGCAGA 205 5789 ACTCGGCTTCGCGCGCAGAG 206 5790 CTCGGCTTCGCGCGCAGAGA 207 5791 TCGGCTTCGCGCGCAGAGAG 208 5792 CGGCTTCGCGCGCAGAGAGC 209 5793 GGCTTCGCGCGCAGAGAGCC 210 5794 GCTTCGCGCGCAGAGAGCCG 211 5795 CTTCGCGCGCAGAGAGCCGA 212 5796 TTCGCGCGCAGAGAGCCGAA 213 5797 TCGCGCGCAGAGAGCCGAAA 214 5798 TGCGCCCCGCGCGGTCCCGT 132 5799 GTGCGCCCCGCGCGGTCCCG 131 5800 GGTGCGCCCCGCGCGGTCCC 130 5801 GGGTGCGCCCCGCGCGGTCC 129 5802 CGGGTGCGCCCCGCGCGGTC 128 5803 GCGGGTGCGCCCCGCGCGGT 127 5804 AGCGGGTGCGCCCCGCGCGG 126 5805 CAGCGGGTGCGCCCCGCGCG 125 5806 TCAGCGGGTGCGCCCCGCGC 124 5807 TTCAGCGGGTGCGCCCCGCG 123 5808 CTTCAGCGGGTGCGCCCCGC 122 5809 CCTTCAGCGGGTGCGCCCCG 121 5810 GCCTTCAGCGGGTGCGCCCC 120 5811 TGCCTTCAGCGGGTGCGCCC 119 5812 CTGCCTTCAGCGGGTGCGCC 118 5813 GCTGCCTTCAGCGGGTGCGC 117 5814 GGCTGCCTTCAGCGGGTGCG 116 5815 GGGCTGCCTTCAGCGGGTGC 115 5816 GGGGCTGCCTTCAGCGGGTG 114 5817 CGGGGCTGCCTTCAGCGGGT 113 5818 CCGGGGCTGCCTTCAGCGGG 112 5819 CCCGGGGCTGCCTTCAGCGG 111 5820 CCCCGGGGCTGCCTTCAGCG 110 5821 GCCCCGGGGCTGCCTTCAGC 109 5822 GGCCCCGGGGCTGCCTTCAG 108 5823 GGGCCCCGGGGCTGCCTTCA 107 5824 CGGGCCCCGGGGCTGCCTTC 106 5825 GCGGGCCCCGGGGCTGCCTT 105 5826 CGCGGGCCCCGGGGCTGCCT 104 5827 CCGCGGGCCCCGGGGCTGCC 103 5828 GCCGCGGGCCCCGGGGCTGC 102 5829 GGCCGCGGGCCCCGGGGCTG 101 5830 GGGCCGCGGGCCCCGGGGCT 100 5831 CGGGCCGCGGGCCCCGGGGC 99 5832 CCGGGCCGCGGGCCCCGGGG 98 5833 TCCGGGCCGCGGGCCCCGGG 97 5834 GTCCGGGCCGCGGGCCCCGG 96 5835 AGTCCGGGCCGCGGGCCCCG 95 5836 AAGTCCGGGCCGCGGGCCCC 94 5837 CAAGTCCGGGCCGCGGGCCC 93 5838 CCAAGTCCGGGCCGCGGGCC 92 5839 ACCAAGTCCGGGCCGCGGGC 91 5840 GACCAAGTCCGGGCCGCGGG 90 5841 GGACCAAGTCCGGGCCGCGG 89 5842 AGGACCAAGTCCGGGCCGCG 88 5843 CAGGACCAAGTCCGGGCCGC 87 5844 GCAGGACCAAGTCCGGGCCG 86 5845 CGCAGGACCAAGTCCGGGCC 85 5846 GCGCAGGACCAAGTCCGGGC 84 5847 TGCGCAGGACCAAGTCCGGG 83 5848 CTGCGCAGGACCAAGTCCGG 82 5849 GCTGCGCAGGACCAAGTCCG 81 5850 CGCTGCGCAGGACCAAGTCC 80 5851 CCGCTGCGCAGGACCAAGTC 79 5852 CCCGCTGCGCAGGACCAAGT 78 5853 GCCCGCTGCGCAGGACCAAG 77 5854 CGCCCGCTGCGCAGGACCAA 76 5855 GCGCCCGCTGCGCAGGACCA 75 5856 CGCGCCCGCTGCGCAGGACC 74 5857 CCGCGCCCGCTGCGCAGGAC 73 5858 CCCGCGCCCGCTGCGCAGGA 72 5859 CCCCGCGCCCGCTGCGCAGG 71 5860 GCCCCGCGCCCGCTGCGCAG 70 5861 TGCCCCGCGCCCGCTGCGCA 69 5862 CTGCCCCGCGCCCGCTGCGC 68 5863 GCTGCCCCGCGCCCGCTGCG 67 5864 CGCTGCCCCGCGCCCGCTGC 66 5865 GCGCTGCCCCGCGCCCGCTG 65 5866 TGCGCTGCCCCGCGCCCGCT 64 5867 CTGCGCTGCCCCGCGCCCGC 63 5868 GCTGCGCTGCCCCGCGCCCG 62 5869 CGCTGCGCTGCCCCGCGCCC 61 5870 CCGCTGCGCTGCCCCGCGCC 60 5871 CCCGCTGCGCTGCCCCGCGC 59 5872 TCCCGCTGCGCTGCCCCGCG 58 5873 CTCCCGCTGCGCTGCCCCGC 57 5874 CCTCCCGCTGCGCTGCCCCG 56 5875 TCCTCCCGCTGCGCTGCCCC 55 5876 TTCCTCCCGCTGCGCTGCCC 54 5877 CTTCCTCCCGCTGCGCTGCC 53 5878 GCTTCCTCCCGCTGCGCTGC 52 5879 CGCTTCCTCCCGCTGCGCTG 51 5880 TCGCTTCCTCCCGCTGCGCT 50 5881 CTCGCTTCCTCCCGCTGCGC 49 5882 TCTCGCTTCCTCCCGCTGCG 48 5883 CTCTCGCTTCCTCCCGCTGC 47 5884 CCTCTCGCTTCCTCCCGCTG 46 5885 ACCTCTCGCTTCCTCCCGCT 45 5886 CACCTCTCGCTTCCTCCCGC 44 5887 GCACCTCTCGCTTCCTCCCG 43 5888 AGCACCTCTCGCTTCCTCCC 42 5889 CAGCACCTCTCGCTTCCTCC 41 5890 GCAGCACCTCTCGCTTCCTC 40 5891 GGCAGCACCTCTCGCTTCCT 39 5892 GGGCAGCACCTCTCGCTTCC 38 5893 AGGGCAGCACCTCTCGCTTC 37 5894 GAGGGCAGCACCTCTCGCTT 36 5895 GGAGGGCAGCACCTCTCGCT 35 5896 GGGAGGGCAGCACCTCTCGC 34 5897 GGGGAGGGCAGCACCTCTCG 33 5898 GCGGAGCGGGCTGAACGTGCG 249 5899 CGGAGCGGGCTGAACGTGCG 250 5900 GACTGGAGGCCGGGGGAGGGGCGGGG 433 5901 GACCCGGGTAACGCGCTTCCAAC 5 5902 ACCCGGGTAACGCGCTTCCA 6 5903 CCCGGGTAACGCGCTTCCAA 7 5904 CCGGGTAACGCGCTTCCAAC 8 5905 CGGGTAACGCGCTTCCAACT 9 5906 GGGTAACGCGCTTCCAACTC 10 5907 GGTAACGCGCTTCCAACTCC 11 5908 GTAACGCGCTTCCAACTCCG 12 5909 TAACGCGCTTCCAACTCCGG 13 5910 AACGCGCTTCCAACTCCGGG 14 5911 ACGCGCTTCCAACTCCGGGG 15 5912 CGCGCTTCCAACTCCGGGGG 16 5913 GCGCTTCCAACTCCGGGGGG 17 5914 CGCTTCCAACTCCGGGGGGA 18 5915 GCTTCCAACTCCGGGGGGAG 19 5916 CTTCCAACTCCGGGGGGAGG 20 5917 TTCCAACTCCGGGGGGAGGG 21 5918 TCCAACTCCGGGGGGAGGGC 22 5919 CCAACTCCGGGGGGAGGGCA 23 5920 GGACCCGGGTAACGCGCTTC 4 5921 TGGACCCGGGTAACGCGCTT 3 5922 TTGGACCCGGGTAACGCGCT 2 5923 TTTGGACCCGGGTAACGCGC 1 5924 CACTCGGCTCCGCCCCTATTGC 507 5925 ACTCGGCTCCGCCCCTATTG 508 5926 CTCGGCTCCGCCCCTATTGC 509 5927 TCGGCTCCGCCCCTATTGCC 510 5928 CGGCTCCGCCCCTATTGCCT 511 5929 GGCTCCGCCCCTATTGCCTC 512 5930 GCTCCGCCCCTATTGCCTCG 513 5931 CTCCGCCCCTATTGCCTCGC 514 5932 TCCGCCCCTATTGCCTCGCA 515 5933 CCGCCCCTATTGCCTCGCAG 516 5934 CGCCCCTATTGCCTCGCAGA 517 5935 GCCCCTATTGCCTCGCAGAC 518 5936 CCCCTATTGCCTCGCAGACA 519 5937 CCCTATTGCCTCGCAGACAA 520 5938 CCTATTGCCTCGCAGACAAC 521 5939 CTATTGCCTCGCAGACAACC 522 5940 TATTGCCTCGCAGACAACCA 523 5941 ATTGCCTCGCAGACAACCAA 524 5942 TTGCCTCGCAGACAACCAAT 525 5943 TGCCTCGCAGACAACCAATG 526 5944 GCCTCGCAGACAACCAATGG 527 5945 CCTCGCAGACAACCAATGGG 528 5946 CTCGCAGACAACCAATGGGG 529 5947 TCGCAGACAACCAATGGGGG 530 5948 CGCAGACAACCAATGGGGGC 531 5949 CCACTCGGCTCCGCCCCTAT 506 5950 CCCACTCGGCTCCGCCCCTA 505 5951 TCCCACTCGGCTCCGCCCCT 504 5952 CTCCCACTCGGCTCCGCCCC 503 5953 ACTCCCACTCGGCTCCGCCC 502 5954 CACTCCCACTCGGCTCCGCC 501 5955 ACACTCCCACTCGGCTCCGC 500 5956 CACACTCCCACTCGGCTCCG 499 5957 CCACACTCCCACTCGGCTCC 498 5958 TCCACACTCCCACTCGGCTC 497 5959 TTCCACACTCCCACTCGGCT 496 5960 TTTCCACACTCCCACTCGGC 495 5961 CTTTCCACACTCCCACTCGG 494 5962 GCTTTCCACACTCCCACTCG 493 5963 CGCTTTCCACACTCCCACTC 492 5964 GCGCTTTCCACACTCCCACT 491 5965 GGCGCTTTCCACACTCCCAC 490 5966 CGGCGCTTTCCACACTCCCA 489 5967 GCGGCGCTTTCCACACTCCC 488 5968 TGCGGCGCTTTCCACACTCC 487 5969 ATGCGGCGCTTTCCACACTC 486 5970 GATGCGGCGCTTTCCACACT 485 5971 GGATGCGGCGCTTTCCACAC 484 5972 GGGATGCGGCGCTTTCCACA 483 5973 CGGGATGCGGCGCTTTCCAC 482 5974 CCGGGATGCGGCGCTTTCCA 481 5975 CCCGGGATGCGGCGCTTTCC 480 5976 ACCCGGGATGCGGCGCTTTC 479 5977 CACCCGGGATGCGGCGCTTT 478 5978 CCACCCGGGATGCGGCGCTT 477 5979 CCCACCCGGGATGCGGCGCT 476 5980 TCCCACCCGGGATGCGGCGC 475 5981 CTCCCACCCGGGATGCGGCG 474 5982 CCTCCCACCCGGGATGCGGC 473 5983 GCCTCCCACCCGGGATGCGG 472 5984 CGCCTCCCACCCGGGATGCG 471 5985 TCGCCTCCCACCCGGGATGC 470 5986 CTCGCCTCCCACCCGGGATG 469 5987 CCTCGCCTCCCACCCGGGAT 468 5988 GCCTCGCCTCCCACCCGGGA 467 5989 AGCCTCGCCTCCCACCCGGG 466 5990 AAGCCTCGCCTCCCACCCGG 465 5991 GAAGCCTCGCCTCCCACCCG 464 5992 GGAAGCCTCGCCTCCCACCC 463 5993 GGGAAGCCTCGCCTCCCACC 462 5994 GGGGAAGCCTCGCCTCCCAC 461 5995 AGGGGAAGCCTCGCCTCCCA 460 5996 AAGGGGAAGCCTCGCCTCCC 459 5997 GAAGGGGAAGCCTCGCCTCC 458 5998 GGAAGGGGAAGCCTCGCCTC 457 5999 GGGAAGGGGAAGCCTCGCCT 456 6000 TACGTCACGGGAGCGCGGCGCAC 578 6001 ACGTCACGGGAGCGCGGCGC 579 6002 CGTCACGGGAGCGCGGCGCA 580 6003 GTCACGGGAGCGCGGCGCAC 581 6004 TCACGGGAGCGCGGCGCACT 582 6005 CACGGGAGCGCGGCGCACTG 583 6006 ACGGGAGCGCGGCGCACTGC 584 6007 CGGGAGCGCGGCGCACTGCC 585 6008 GGGAGCGCGGCGCACTGCCT 586 6009 GGAGCGCGGCGCACTGCCTG 587 6010 GAGCGCGGCGCACTGCCTGG 588 6011 AGCGCGGCGCACTGCCTGGG 589 6012 GCGCGGCGCACTGCCTGGGG 590 6013 CGCGGCGCACTGCCTGGGGG 591 6014 GCGGCGCACTGCCTGGGGGC 592 6015 CGGCGCACTGCCTGGGGGCG 593 6016 GGCGCACTGCCTGGGGGCGG 594 6017 GCGCACTGCCTGGGGGCGGG 595 6018 CGCACTGCCTGGGGGCGGGG 596 6019 GCACTGCCTGGGGGCGGGGT 597 6020 CACTGCCTGGGGGCGGGGTC 598 6021 ACTGCCTGGGGGCGGGGTCC 599 6022 CTGCCTGGGGGCGGGGTCCG 600 6023 TGCCTGGGGGCGGGGTCCGT 601 6024 GCCTGGGGGCGGGGTCCGTC 602 6025 CCTGGGGGCGGGGTCCGTCG 603 6026 CTGGGGGCGGGGTCCGTCGC 604 6027 TGGGGGCGGGGTCCGTCGCG 605 6028 GGGGGCGGGGTCCGTCGCGG 606 6029 GGGGCGGGGTCCGTCGCGGA 607 6030 GGGCGGGGTCCGTCGCGGAC 608 6031 GGCGGGGTCCGTCGCGGACG 609 6032 GCGGGGTCCGTCGCGGACGC 610 6033 CGGGGTCCGTCGCGGACGCC 611 6034 GGGGTCCGTCGCGGACGCCG 612 6035 GGGTCCGTCGCGGACGCCGT 613 6036 GGTCCGTCGCGGACGCCGTG 614 6037 GTCCGTCGCGGACGCCGTGG 615 6038 TCCGTCGCGGACGCCGTGGC 616 6039 CCGTCGCGGACGCCGTGGCG 617 6040 CGTCGCGGACGCCGTGGCGC 618 6041 GTCGCGGACGCCGTGGCGCC 619 6042 TCGCGGACGCCGTGGCGCCC 620 6043 CGCGGACGCCGTGGCGCCCT 621 6044 GCGGACGCCGTGGCGCCCTC 622 6045 CGGACGCCGTGGCGCCCTCT 623 6046 GGACGCCGTGGCGCCCTCTG 624 6047 GACGCCGTGGCGCCCTCTGT 625 6048 ACGCCGTGGCGCCCTCTGTC 626 6049 CGCCGTGGCGCCCTCTGTCG 627 6050 GCCGTGGCGCCCTCTGTCGC 628 6051 CCGTGGCGCCCTCTGTCGCC 629 6052 CGTGGCGCCCTCTGTCGCCC 630 6053 GTGGCGCCCTCTGTCGCCCC 631 6054 TGGCGCCCTCTGTCGCCCCG 632 6055 GGCGCCCTCTGTCGCCCCGA 633 6056 GCGCCCTCTGTCGCCCCGAG 634 6057 CGCCCTCTGTCGCCCCGAGG 635 6058 GCCCTCTGTCGCCCCGAGGC 636 6059 CCCTCTGTCGCCCCGAGGCA 637 6060 CCTCTGTCGCCCCGAGGCAA 638 6061 CTCTGTCGCCCCGAGGCAAG 639 6062 TCTGTCGCCCCGAGGCAAGC 640 6063 CTGTCGCCCCGAGGCAAGCA 641 6064 TGTCGCCCCGAGGCAAGCAG 642 6065 GTCGCCCCGAGGCAAGCAGG 643 6066 TCGCCCCGAGGCAAGCAGGT 644 6067 CGCCCCGAGGCAAGCAGGTG 645 6068 GCCCCGAGGCAAGCAGGTGG 646 6069 CCCCGAGGCAAGCAGGTGGA 647 6070 CCCGAGGCAAGCAGGTGGAC 648 6071 CCGAGGCAAGCAGGTGGACC 649 6072 CGAGGCAAGCAGGTGGACCC 650 6073 ATACGTCACGGGAGCGCGGC 577 6074 AATACGTCACGGGAGCGCGG 576 6075 AAATACGTCACGGGAGCGCG 575 6076 GAAATACGTCACGGGAGCGC 574 6077 GTCGCGGACGCCGTGGCGCCCTCTGTC 619 6078 TCGCGGACGCCGTGGCGCCC 620 6079 CGCGGACGCCGTGGCGCCCT 621 6080 GCGGACGCCGTGGCGCCCTC 622 6081 CGGACGCCGTGGCGCCCTCT 623 6082 GGACGCCGTGGCGCCCTCTG 624 6083 GACGCCGTGGCGCCCTCTGT 625 6084 ACGCCGTGGCGCCCTCTGTC 626 6085 CGCCGTGGCGCCCTCTGTCG 627 6086 GCCGTGGCGCCCTCTGTCGC 628 6087 CCGTGGCGCCCTCTGTCGCC 629 6088 CGTGGCGCCCTCTGTCGCCC 630 6089 GTGGCGCCCTCTGTCGCCCC 631 6090 TGGCGCCCTCTGTCGCCCCG 632 6091 GGCGCCCTCTGTCGCCCCGA 633 6092 GCGCCCTCTGTCGCCCCGAG 634 6093 CGCCCTCTGTCGCCCCGAGG 635 6094 GCCCTCTGTCGCCCCGAGGC 636 6095 CCCTCTGTCGCCCCGAGGCA 637 6096 CCTCTGTCGCCCCGAGGCAA 638 6097 CTCTGTCGCCCCGAGGCAAG 639 6098 TCTGTCGCCCCGAGGCAAGC 640 6099 CTGTCGCCCCGAGGCAAGCA 641 6100 TGTCGCCCCGAGGCAAGCAG 642 6101 GTCGCCCCGAGGCAAGCAGG 643 6102 TCGCCCCGAGGCAAGCAGGT 644 6103 CGCCCCGAGGCAAGCAGGTG 645 6104 GCCCCGAGGCAAGCAGGTGG 646 6105 CCCCGAGGCAAGCAGGTGGA 647 6106 CCCGAGGCAAGCAGGTGGAC 648 6107 CCGAGGCAAGCAGGTGGACC 649 6108 CGAGGCAAGCAGGTGGACCC 650 6109 CGTCGCGGACGCCGTGGCGC 618 6110 CCGTCGCGGACGCCGTGGCG 617 6111 TCCGTCGCGGACGCCGTGGC 616 6112 GTCCGTCGCGGACGCCGTGG 615 6113 GGTCCGTCGCGGACGCCGTG 614 6114 GGGTCCGTCGCGGACGCCGT 613 6115 GGGGTCCGTCGCGGACGCCG 612 6116 CGGGGTCCGTCGCGGACGCC 611 6117 GCGGGGTCCGTCGCGGACGC 610 6118 GGCGGGGTCCGTCGCGGACG 609 6119 GGGCGGGGTCCGTCGCGGAC 608 6120 GGGGCGGGGTCCGTCGCGGA 607 6121 GGGGGCGGGGTCCGTCGCGG 606 6122 TGGGGGCGGGGTCCGTCGCG 605 6123 CTGGGGGCGGGGTCCGTCGC 604 6124 CCTGGGGGCGGGGTCCGTCG 603 6125 GCCTGGGGGCGGGGTCCGTC 602 6126 TGCCTGGGGGCGGGGTCCGT 601 6127 CTGCCTGGGGGCGGGGTCCG 600 6128 ACTGCCTGGGGGCGGGGTCC 599 6129 CACTGCCTGGGGGCGGGGTC 598 6130 GCACTGCCTGGGGGCGGGGT 597 6131 CGCACTGCCTGGGGGCGGGG 596 6132 GCGCACTGCCTGGGGGCGGG 595 6133 GGCGCACTGCCTGGGGGCGG 594 6134 CGGCGCACTGCCTGGGGGCG 593 6135 GCGGCGCACTGCCTGGGGGC 592 6136 CGCGGCGCACTGCCTGGGGG 591 6137 GCGCGGCGCACTGCCTGGGG 590 6138 AGCGCGGCGCACTGCCTGGG 589 6139 GAGCGCGGCGCACTGCCTGG 588 6140 GGAGCGCGGCGCACTGCCTG 587 6141 GGGAGCGCGGCGCACTGCCT 586 6142 CGGGAGCGCGGCGCACTGCC 585 6143 ACGGGAGCGCGGCGCACTGC 584 6144 CACGGGAGCGCGGCGCACTG 583 6145 TCACGGGAGCGCGGCGCACT 582 6146 GTCACGGGAGCGCGGCGCAC 581 6147 CGTCACGGGAGCGCGGCGCA 580 6148 ACGTCACGGGAGCGCGGCGC 579 6149 TACGTCACGGGAGCGCGGCG 578 6150 ATACGTCACGGGAGCGCGGC 577 6151 AATACGTCACGGGAGCGCGG 576 6152 AAATACGTCACGGGAGCGCG 575 6153 GAAATACGTCACGGGAGCGC 574 6154 CACTCGCCGTCATGCCCGGATCC 1183 6155 ACTCGCCGTCATGCCCGGAT 1184 6156 CTCGCCGTCATGCCCGGATC 1185 6157 TCGCCGTCATGCCCGGATCC 1186 6158 CGCCGTCATGCCCGGATCCT 1187 6159 GCCGTCATGCCCGGATCCTT 1188 6160 CCGTCATGCCCGGATCCTTT 1189 6161 CGTCATGCCCGGATCCTTTT 1190 6162 GTCATGCCCGGATCCTTTTT 1191 6163 TCATGCCCGGATCCTTTTTG 1192 6164 CATGCCCGGATCCTTTTTGT 1193 6165 ATGCCCGGATCCTTTTTGTA 1194 6166 TGCCCGGATCCTTTTTGTAT 1195 6167 GCCCGGATCCTTTTTGTATT 1196 6168 GCACTCGCCGTCATGCCCGG 1182 6169 GGCACTCGCCGTCATGCCCG 1181 6170 AGGCACTCGCCGTCATGCCC 1180 6171 CAGGCACTCGCCGTCATGCC 1179 6172 ACAGGCACTCGCCGTCATGC 1178 6173 TACAGGCACTCGCCGTCATG 1177 6174 TTACAGGCACTCGCCGTCAT 1176 6175 ATTACAGGCACTCGCCGTCA 1175 6176 GATTACAGGCACTCGCCGTC 1174 6177 GGATTACAGGCACTCGCCGT 1173 6178 GGGATTACAGGCACTCGCCG 1172 6179 TGGGATTACAGGCACTCGCC 1171 6180 CTGGGATTACAGGCACTCGC 1170 6181 GCTGGGATTACAGGCACTCG 1169

Hot Zones (Relative upstream location to gene start site)  1-950 1050-1500

Examples

In FIG. 37, In HCT-116 (human colorectal carcinoma), MEK1_(—)1 (216) and MEK1_(—)2 (212) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The MEK1 sequences MEK1_(—)1 (216) and MEK1_(—)2 (212) fit the independent and dependent DNAi motif claims.

The secondary structures for MEK1_(—)1 (216) and MEK1_(—)2 (212) are shown in FIGS. 38 and 39.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11969) ACATATAGTTCAGTCTTATTCTTGTCTGTATGGTCAGCACTTATGTTAGG CCCTCAGGAAAAGTTGACAGAACCGATGGATCACTGCCGGTCTGAAAAGG AAATGAGGAAAACAAATTCTCCTACCTTGAACTATTCTGCAAACTTTAAC CATTGGGGTAATTGTTTATCTGGGCTTCTTGGATCATGATAAGGGCTTAG GGTTTACTCAGTGGAGGCCAACCCAGCATGCATAGAATCATAATATTTCA ATATTAAAAAGAATGCTGCATTTTACACAGAGTGGAAGTGAGGCCTTGAA AATTTCAATTAATTGCTCAAAGTCCTAATAGTTTTTATTTGAACTAGTAA ATATAAAATTATACCAGAATTCAGATAGACTGCCTTGATAATAGATTACT TTGAAAAGTTTCAATTTTTTTTTTTTTTTTGAGATAGTCTCACTGTGTTG CACAGGCTGGAGTACAGTGGAGTGATCTTGGCTCACTGTAACCTCCACCT CCTGGGTTCAAGTGATTCTCCAGCTTCAGCCTCCCAAGTAGCTGGGACTA CAGGCACCCGCCACCACATTCAGATAATTTTTGTATTTTTAGTAAAGACA GGGTTTCACCATGTTGGCCAGGCTTGGTCTTGAACTCCTGACCTCAGGTG ATCCTCCCACCTCAGCCTCCCAAAGTGCTGGGATTAAAGGTGTGAGCCAC CACCACACCTGGCCTTCAATTCACTTTTTAATGTTTATTATTTTACTCTG ATACTAAAAATTATGCATGTTTAACATGAATAAGGACACACTTCTACACA CACATGCATACATTTACATCTATGCCTCTATATTAAAAAGTATGGGGGAA AGAAATGGGGAGATGTAGGTCAAAGAATATAAAGCAGCAGATATGTAGGA TGAAGAAGTCTAGAGATCTAATGTACAACATGAAGACCATAGTTAATAAC ATTGTATTTTATTTGCGTTTTTTGTTAAATAAGTAGATTTTAGCTGCTCG TCATACTTTACACAAGCCTTTATGTGACGGTATAGATATGTTAATTCACT TCACTATAGTAACCATTTTACTATCTATATATATCCCATAACATCATGTT ACAAACCTCAAATATACACAATAAAATTTATTTTTATTTATTTAATTTAT TTATTTATTTTTGAGACGGAGTCTTGTTCTGTCGCCCAGGCTGGAGTGCA GTGGCGCGATCTCGGCTCACTGCAAGCTCCACCTCCCGGGTTCACACCAT TCTCCTGCCTCAGCCTCCTGAGTAGCTGGGACTACAGGCACCCACCACCA CGCCCGGCTAATTTTTTGTATTTTTTAGTAGAGATGGGGTTTCACCGTGT TAGCCAGGATGGTCTCGATTTCCTGACCTCGTGATCTGCCCACCTCAGCC TCCCAAAGTGCTGGGATTACAGGCATGAGCCACCGCGCCCGGCCTATTTT ATTTATTTTTGAGACAGAGTCTTGCTCTGTTGCCCAGGCTGGAGTGCAGT GGTGCAATCTCGGCTCACTGCAAACTCTGCCTCCCTGGTTCAGGCAATTA TCCTGCCTCAGCCTCCTGAGTAGCTGGGATTACAGGTGCCCACCACCATG CCTGGCTAATTTTTGTAATTTAGTAGAGACGGGGTTTCACCATGTTGGCC AGGCTGATCTTGAAGTCCTGACCTCAAGTGATCTTCCAGCTTTGGCCTCA CAAAGTGCTGGGATTACAGGTGGTAGCCGCCACTGCATCCACCCAGAATA ATTTATTTTTTAAAAAACTATGAGTTCAGGCCGGGCGCAGTGGCTCACGC CTGTAAACCCAGCACTTTGGGAGGCCGAGGTGGGCGGATCACCTGAGGTC AGGAGTTTGAGACCAGCCTGGCCAACATGGTGAAATCCTGTCTCTACTAA AAATACAAAATTAGCCAGGCATGGTGGTGCATGCCTGTAATCCCAGCTAC TTGGGAGGCTGAGGCAGGAGAATCACTTGAGCTTGGGAGGTGGAGGTTGC AATGAGCCAAGGTTGCGCCATTGCACTCAAGCCTGGGCAAAAAGAGCAAA ACGCCACTCAAAAACAAAAACAAAACAAAACAAAAACACCCCCCCAAAAA ACAAAACAAAACAATGAGTTCACACTGATACCTCCAATTCCAATACAATA GCGTAAGGTATTCTCCCTTCCCATACTTCTAACGTCATTCTACCACAGTG AGAAAGCTGGCTCTGTCATGCTTAATATATTTAGTGACTTAATCAACCAT CCTGAATGCAACTAACCTCCCATCTAAGCTTCTAGGCCTTCCCCACTTGG ATGCCTTGTTCTCCCCTCTTGGGCCCTACGGCTAAGACTTTGTGTAGGAC TGCCTCCCAGGTGTTCAAGCCCTCTTCATTTTCTCAGGTTCCTCAGCCTC CTTACCTGCTAGGTCACCAACACCTGGCTGTGGATAACCAGGTGTAGATG TTTCCTTTGTTCTGTACACGTTTCCTTTGTTCTGTACACCTAATGTCTTT GACACTTAGTATTTTAGGATGGGAAAGGGGAAGAGGAACACTGAATGTGC ACTTTTAAATGGGTATTGTGCCTCTTATTAAGCTCTTTATTCACATCTTA TTTCTTTAGTAATTCACAGAATTGGAATTTTTGGATTAAAGTTCTTTTTT TTTTTGAGACGGGGTCTCACTCTGTCGCCCAGGCTGGAGTGCAGTGGTGT GATCTTGGATCACTGCAACCTCCGCCTCCCGAGTTCAAGCAATTCTCTGC CTCAGCCCCCCAAGTAGTTGGGATTACAGGCACCCGCCACCACGCCCAGC TAATTTTTTGTATTTTTAGTAGAGATGGGTTTCACCATCTTGGCCAGGCT GGTCTTGAACTCCTGACCTCGTGATCCACCCGTCTCGGCCTCCCAAAGTT CTGGAATTACAGGCGTGAGCCACCGCGCCTGGCCTGGATGAAAGTTTTTT TAAAGGGAGTCTTGCTCTGTAGCCCTGGCTGGTGTGCAGTGGTGTGATCA TAGCTCACTGCAGCCTCAAACTCCTGGGCTCAAGTGATCCTCCAGCCTCA GCCTCCTCAGTAGCTTGGACGACAGCTGCACACAACCATGCCCAGCTAAT AGAGACGGGGGACTCACTATGTTGCCCAGGCTAGTCTCGAACTCCTGGGC TCAAGTGATCCTCTTGCCTGGGCCTCCCAAAATTGGGATTACAGGCGTGA GCCACCGCTCCTGGCCCGAAAGAGTGTTTTTAAGGCTTTAAAAAAATATT GCCAACATGGTGAAAACCCGTTTCTACAAAAATACAAAAAGGATCCGGGC ATGACGGCGAGTGCCTGTAATCCCAGCTACTCAGGAGACTGAGGCAGGAG AATCGCTTGAACGTGGGAGGCAGAGGTGGTAGTTAGCGGAGATCGCGCCA CTACACTCCAGGCTGGGCAACTGAGGGAGACACCGTCTTAAAAAAAAAAA AGTTCCCAAGTCTAAAAAAAAAAAAATCATCAATCTGCTCTCAAAAACTG TCGCAACAATTTACAATCTCATCAGCACTGAGTATCCATTTCCTTGCACC CTTCTCAGTAGTATTACCATTAAACAAACAAAATTTATATGCGTCAGTTT GTTGGGCTCAAAGGAGCCTCTCGACAAGTTTCCTATTCCCCACGCTGCCT CTCCTCTGGACACAGGAAGGGGTCCTTTTCCTTATTTATTTTGTTATTTC ATTTTCGTCAACACGGCTCGGCTTGGGGACAGGGGTCGGGGGCAGGCCGG TTACCGCAGAGGTGGAGGCCGCGCGGCACCTGGCCTGGAGAGCTCACCAC ACAGCGACACAGACTTCTTCTCAGCTGGGTCCACCTGCTTGCCTCGGGGC GACAGAGGGCGCCACGGCGTCCGCGACGGACCCCGCCCCCAGGCAGTGCG CCGCGCTCCCGTGACGTATTTCCGCGTCATCTGCCGCCGAGGCTTGCCCC CATTGGTTGTCTGCGAGGCAATAGGGGCGGAGCCGAGTGGGAGTGTGGAA AGCGCCGCATCCCGGGTGGGAGGCGAGGCTTCCCCTTCCCCGCCCCTCCC CCGGCCTCCAGTCCCTCCCAGGGCCGCTTCGCAGAGCGGCTAGGAGCACG GCGGCGGCGGCACTTTCCCCGGCAGGAGCTGGAGCTGGGCTCTGGTGCGC GCGCGGCTGTGCCGCCCGAGCCGGAGGGACTGGTTGGTTGAGAGAGAGAG AGGAAGGGAATCCCGGGCTGCCGAACCGCACGTTCAGCCCGCTCCGCTCC TGCAGGGCAGCCTTTCGGCTCTCTGCGCGCGAAGCCGAGTCCCGGGCGGG TGGGGCGGGGGTCCACTGAGACCGCTACCGGCCCCTCGGCGCTGACGGGA CCGCGCGGGGCGCACCCGCTGAAGGCAGCCCCGGGGCCCGCGGCCCGGAC TTGGTCCTGCGCAGCGGGCGCGGGGCAGCGCAGCGGGAGGAAGCGAGAGG TGCTGCCCTCCCCCCGGAGTTGGAAGCGCGTTACCCGGGTCCAAA ATG

21) MEK1 and MEK2 (MAP2K2) Mitogen-activated protein kinase kinase 1. Dual specificity protein kinases act as an essential component of the MAP kinase signal transduction pathway and serves as an integration point for multiple biochemical signals. MEK1 and MEK2 are members of the dual specificity protein kinase family, which act as a mitogen-activated protein (MAP) kinase kinases and as extracellular signal-regulated kinases (ERKs). Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. RAF1 then further activates the dual-specificity protein kinases MAP2K1/MEK1 and MAP2K2/MEK2. Both MAP2K1/MEK1 and MAP2K2/MEK2 function specifically in the MAPK/ERK cascade, and catalyze the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, leading to their activation and further transduction of the signal within the MAPK/ERK cascade. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth and proliferation, adhesion, survival and differentiation, predominantly through the regulation of transcription, metabolism and cytoskeletal rearrangements (reviewed by Roberts and Der; 2007 Oncogene 26, 3291-3310).

Genetic alterations that activate the mitogen-activated protein kinase (MAP kinase) pathway occur commonly in cancer. For example, the majority of melanomas harbor mutations in the BRAF oncogene, which confers enhanced sensitivity to pharmacologic MAP kinase inhibition (e.g., RAF or MEK inhibitors). Most mutations conferring resistance to MEK inhibition in vitro populated the allosteric drug binding pocket or alpha-helix C and showed robust (approximately 100-fold) resistance to allosteric MEK inhibition (reviewed in Emery et al, 2009; Proc Natl Acad Sci.; 106(48):20411-20416). Other mutations affected MEK1 codons located within or abutting the N-terminal negative regulatory helix (helix A), which also undergo gain-of-function germline mutations in cardiofaciocutaneous (CFC) syndrome. One target of the MAPK/ERK cascade is peroxisome proliferator-activated receptor gamma (PPARG), a nuclear receptor that promotes differentiation and apoptosis. MAP2K1/MEK1 has been shown to export PPARG from the nucleus. The MAPK/ERK cascade is also involved in the regulation of endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC), as well as in the fragmentation of the Golgi apparatus during mitosis.

Protein: MEK2 Gene: MAP2K2 (Homo sapiens, chromosome 19, 4090319-4124126 [NCBI Reference Sequence: NC_(—)000019.9]; start site location: 4123872; strand: negative)

Gene Identification GeneID 5605 HGNC 6842 HPRD 03164 MIM 601263

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 6182 CGCCGCAGCCCGAGTCCGAGAGG 226 6202 GAGGGGCGCTGGGGCTGAGGCGAGCG 165 6203 CTCGCGATAACGGGATCGGGAGCCGCG 290 6235 MEK2_1 CCGACGCGAGGCGGTGCCGGGACCGG 391 6240 CACGGCGCGTGTGCCCAAGCGC 436 6299 CGTGGACACACGCCCCTAGCCC 643 6341 TAGACACTTCGGTGAATCGTGCCGC 1622

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 6182 CGCCGCAGCCCGAGTCCGAGAGG 226 6183 GCCGCAGCCCGAGTCCGAGA 227 6184 CCGCAGCCCGAGTCCGAGAG 228 6185 CGCAGCCCGAGTCCGAGAGG 229 6186 GCAGCCCGAGTCCGAGAGGC 230 6187 CAGCCCGAGTCCGAGAGGCA 231 6188 AGCCCGAGTCCGAGAGGCAG 232 6189 GCCCGAGTCCGAGAGGCAGG 233 6190 CCCGAGTCCGAGAGGCAGGG 234 6191 ACGCCGCAGCCCGAGTCCGA 225 6192 GACGCCGCAGCCCGAGTCCG 224 6193 TGACGCCGCAGCCCGAGTCC 223 6194 CTGACGCCGCAGCCCGAGTC 222 6195 GCTGACGCCGCAGCCCGAGT 221 6196 GGCTGACGCCGCAGCCCGAG 220 6197 AGGCTGACGCCGCAGCCCGA 219 6198 AAGGCTGACGCCGCAGCCCG 218 6199 GAAGGCTGACGCCGCAGCCC 217 6200 AGAAGGCTGACGCCGCAGCC 216 6201 AAGAAGGCTGACGCCGCAGC 215 6202 GAGGGGCGCTGGGGCTGAGGCGAGCG 165 6203 CTCGCGATAACGGGATCGGGAGCCGCG 291 6204 TCGCGATAACGGGATCGGGA 292 6205 TCTCGCGATAACGGGATCGG 290 6206 TTCTCGCGATAACGGGATCG 289 6207 CTTCTCGCGATAACGGGATC 288 6208 GCTTCTCGCGATAACGGGAT 287 6209 GGCTTCTCGCGATAACGGGA 286 6210 CGGCTTCTCGCGATAACGGG 285 6211 CCGGCTTCTCGCGATAACGG 284 6212 ACCGGCTTCTCGCGATAACG 283 6213 GACCGGCTTCTCGCGATAAC 282 6214 GGACCGGCTTCTCGCGATAA 281 6215 CGGACCGGCTTCTCGCGATA 280 6216 GCGGACCGGCTTCTCGCGAT 279 6217 CGCGGACCGGCTTCTCGCGA 278 6218 TCGCGGACCGGCTTCTCGCG 277 6219 ATCGCGGACCGGCTTCTCGC 276 6220 GATCGCGGACCGGCTTCTCG 275 6221 AGATCGCGGACCGGCTTCTC 274 6222 AAGATCGCGGACCGGCTTCT 273 6223 CAAGATCGCGGACCGGCTTC 272 6224 ACAAGATCGCGGACCGGCTT 271 6225 CACAAGATCGCGGACCGGCT 270 6226 CCACAAGATCGCGGACCGGC 269 6227 GCCACAAGATCGCGGACCGG 268 6228 GGCCACAAGATCGCGGACCG 267 6229 CGGCCACAAGATCGCGGACC 266 6230 GCGGCCACAAGATCGCGGAC 265 6231 GGCGGCCACAAGATCGCGGA 264 6232 GGGCGGCCACAAGATCGCGG 263 6233 GGGGCGGCCACAAGATCGCG 262 6234 AGGGGCGGCCACAAGATCGC 261 6235 CCGACGCGAGGCGGTGCCGGGACCGG 391 6236 CGACGCGAGGCGGTGCCGGG 392 6237 ACCGACGCGAGGCGGTGCCG 390 6238 GACCGACGCGAGGCGGTGCC 389 6239 AGACCGACGCGAGGCGGTGC 388 6240 CACGGCGCGTGTGCCCAAGCGC 436 6241 ACGGCGCGTGTGCCCAAGCG 437 6242 CGGCGCGTGTGCCCAAGCGC 438 6243 GGCGCGTGTGCCCAAGCGCT 439 6244 GCGCGTGTGCCCAAGCGCTT 440 6245 CGCGTGTGCCCAAGCGCTTG 441 6246 GCGTGTGCCCAAGCGCTTGG 442 6247 CGTGTGCCCAAGCGCTTGGG 443 6248 GTGTGCCCAAGCGCTTGGGG 444 6249 TGTGCCCAAGCGCTTGGGGC 445 6250 GTGCCCAAGCGCTTGGGGCA 446 6251 TGCCCAAGCGCTTGGGGCAT 447 6252 GCCCAAGCGCTTGGGGCATG 448 6253 CCCAAGCGCTTGGGGCATGA 449 6254 CCAAGCGCTTGGGGCATGAG 450 6255 CAAGCGCTTGGGGCATGAGG 451 6256 AAGCGCTTGGGGCATGAGGC 452 6257 AGCGCTTGGGGCATGAGGCG 453 6258 GCGCTTGGGGCATGAGGCGC 454 6259 CGCTTGGGGCATGAGGCGCG 455 6260 GCTTGGGGCATGAGGCGCGG 456 6261 CTTGGGGCATGAGGCGCGGG 457 6262 CCACGGCGCGTGTGCCCAAG 435 6263 ACCACGGCGCGTGTGCCCAA 434 6264 TACCACGGCGCGTGTGCCCA 433 6265 TTACCACGGCGCGTGTGCCC 432 6266 CTTACCACGGCGCGTGTGCC 431 6267 CCTTACCACGGCGCGTGTGC 430 6268 GCCTTACCACGGCGCGTGTG 429 6269 TGCCTTACCACGGCGCGTGT 428 6270 TTGCCTTACCACGGCGCGTG 427 6271 CTTGCCTTACCACGGCGCGT 426 6272 GCTTGCCTTACCACGGCGCG 425 6273 CGCTTGCCTTACCACGGCGC 424 6274 TCGCTTGCCTTACCACGGCG 423 6275 CTCGCTTGCCTTACCACGGC 422 6276 CCTCGCTTGCCTTACCACGG 421 6277 CCCTCGCTTGCCTTACCACG 420 6278 GCCCTCGCTTGCCTTACCAC 419 6279 CGCCCTCGCTTGCCTTACCA 418 6280 GCGCCCTCGCTTGCCTTACC 417 6281 GGCGCCCTCGCTTGCCTTAC 416 6282 GGGCGCCCTCGCTTGCCTTA 415 6283 CGGGCGCCCTCGCTTGCCTT 414 6284 CCGGGCGCCCTCGCTTGCCT 413 6285 ACCGGGCGCCCTCGCTTGCC 412 6286 GACCGGGCGCCCTCGCTTGC 411 6287 GGACCGGGCGCCCTCGCTTG 410 6288 GGGACCGGGCGCCCTCGCTT 409 6289 CGGGACCGGGCGCCCTCGCT 408 6290 CCGGGACCGGGCGCCCTCGC 407 6291 GCCGGGACCGGGCGCCCTCG 406 6292 TGCCGGGACCGGGCGCCCTC 405 6293 GTGCCGGGACCGGGCGCCCT 404 6294 GGTGCCGGGACCGGGCGCCC 403 6295 CGGTGCCGGGACCGGGCGCC 402 6296 GCGGTGCCGGGACCGGGCGC 401 6297 GGCGGTGCCGGGACCGGGCG 400 6298 AGGCGGTGCCGGGACCGGGC 399 6299 CGTGGACACACGCCCCTAGCCC 648 6300 GTGGACACACGCCCCTAGCC 649 6301 TGGACACACGCCCCTAGCCC 650 6302 GGACACACGCCCCTAGCCCC 651 6303 GACACACGCCCCTAGCCCCC 652 6304 ACACACGCCCCTAGCCCCCA 653 6305 CACACGCCCCTAGCCCCCAC 654 6306 ACACGCCCCTAGCCCCCACC 655 6307 CACGCCCCTAGCCCCCACCG 656 6308 ACGCCCCTAGCCCCCACCGC 657 6309 CGCCCCTAGCCCCCACCGCC 658 6310 GCCCCTAGCCCCCACCGCCT 659 6311 CCCCTAGCCCCCACCGCCTT 660 6312 CCCTAGCCCCCACCGCCTTA 661 6313 CCTAGCCCCCACCGCCTTAG 662 6314 CTAGCCCCCACCGCCTTAGA 663 6315 TAGCCCCCACCGCCTTAGAG 664 6316 AGCCCCCACCGCCTTAGAGT 665 6317 GCCCCCACCGCCTTAGAGTG 666 6318 CCCCCACCGCCTTAGAGTGT 667 6319 CCCCACCGCCTTAGAGTGTC 668 6320 CCCACCGCCTTAGAGTGTCA 669 6321 CCACCGCCTTAGAGTGTCAG 670 6322 CACCGCCTTAGAGTGTCAGT 671 6323 ACCGCCTTAGAGTGTCAGTT 672 6324 CCGCCTTAGAGTGTCAGTTA 673 6325 CGCCTTAGAGTGTCAGTTAC 674 6326 GCGTGGACACACGCCCCTAG 647 6327 AGCGTGGACACACGCCCCTA 646 6328 AAGCGTGGACACACGCCCCT 645 6329 CAAGCGTGGACACACGCCCC 644 6330 GCAAGCGTGGACACACGCCC 643 6331 GGCAAGCGTGGACACACGCC 642 6332 TGGCAAGCGTGGACACACGC 641 6333 TTGGCAAGCGTGGACACACG 640 6334 TTTGGCAAGCGTGGACACAC 639 6335 TTTTGGCAAGCGTGGACACA 638 6336 TTTTTGGCAAGCGTGGACAC 637 6337 CTTTTTGGCAAGCGTGGACA 636 6338 TCTTTTTGGCAAGCGTGGAC 635 6339 ATCTTTTTGGCAAGCGTGGA 634 6340 AATCTTTTTGGCAAGCGTGG 633 6341 TAGACACTTCGGTGAATCGTGCCGC 1598 6342 AGACACTTCGGTGAATCGTG 1599 6343 GACACTTCGGTGAATCGTGC 1600 6344 ACACTTCGGTGAATCGTGCC 1601 6345 CACTTCGGTGAATCGTGCCG 1602 6346 ACTTCGGTGAATCGTGCCGC 1603 6347 CTTCGGTGAATCGTGCCGCT 1604 6348 TTCGGTGAATCGTGCCGCTA 1605 6349 TCGGTGAATCGTGCCGCTAT 1606 6350 CGGTGAATCGTGCCGCTATG 1607 6351 GGTGAATCGTGCCGCTATGA 1608 6352 GTGAATCGTGCCGCTATGAA 1609 6353 TGAATCGTGCCGCTATGAAC 1610 6354 GAATCGTGCCGCTATGAACA 1611 6355 AATCGTGCCGCTATGAACAC 1612 6356 ATCGTGCCGCTATGAACACA 1613 6357 TCGTGCCGCTATGAACACAG 1614 6358 CGTGCCGCTATGAACACAGA 1615 6359 GTGCCGCTATGAACACAGAT 1616 6360 TGCCGCTATGAACACAGATG 1617 6361 GCCGCTATGAACACAGATGT 1618 6362 CCGCTATGAACACAGATGTA 1619 6363 CGCTATGAACACAGATGTAC 1620 6364 CTAGACACTTCGGTGAATCG 1597 6365 ACTAGACACTTCGGTGAATC 1596 6366 CACTAGACACTTCGGTGAAT 1595 6367 CCACTAGACACTTCGGTGAA 1594 6368 GCCACTAGACACTTCGGTGA 1593 6369 TGCCACTAGACACTTCGGTG 1592 6370 CTGCCACTAGACACTTCGGT 1591 6371 TCTGCCACTAGACACTTCGG 1590 6372 ATCTGCCACTAGACACTTCG 1589

Hot Zones (Relative upstream location to gene start site)  1-750  900-1700 2550-2900 4150-4500

Examples

In FIG. 40, In HCT-116 (human colorectal carcinoma cell line), MEK2_(—)1 (224) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The MEK2 sequences MEK2_(—)1 (224) fits the independent and dependent DNAi motif claims.

The secondary structure for MEK2_(—)1 (224) is shown in FIG. 41.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11970) GGAACTACAGGTGCCCGCCACCACGCCTGGCTAATTTTTTTGTATTTTTA GTAGAGACAGGGTTTCACTGTGTTAGCCAGGATGGTCTCTGGTCTCGATC TCCTGACCTCGTGATCTGCCTGCCTCAGCCTCCCAAAGTGCTGGGATTAC AGGCGTGAGCCACCGCGCCCGGCCTTGTATTTTTAGTAGAGACAGGGTTT GTCCATGTTGGTCAGGCTGGTATCGAACTCCCGACCTCAGGTGATCCACC CGCCTCGGCCTCCCAAAGTGCAGGGATTATAGGCATGAGCCACCACATCT GGTCTTCTTCTTTTTTTTTTTTTTTTTGAGACAGAGTCTCCCTCAGGCTG GAGTGCGGTGGCACGATCTTGGCTCACTGCAACCTCCACCTCTCAGGTTC AAGTAATTCTCGTGCCTCAGCCTCCCAAGTAGCTGAGACTACAGGCACCT GCCACCATGCCCAGCTAATTTTTTTTTTTTTTCCGAGATGGAGCCTTACT CTGTTGCCCAGGCTGGAGTGCAGGGGCACAATCTTGGCTCACTGCAACCT CCACCTCCGGGGTTCAAGCAGTTCTCCTGCCTCAGCCTCCCGAGTAGCTG GGATTACAGGTGCCCACCACCATGCCCGGCTAATTTTTGTGTGTTTTTAG TAGAGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTTGAACTCTTGACC TCAGGTGATCTGCCCACCTCGGCTTGCCAAAGTGCTGTGATTACACCCGT GACCAGCCTAATTTTTGTATATTTAGTAGAGATGGGGTTTCACCATGTTG GCCAGGCTGGACTCGAACTCCTGACCTCAAGTGATCACCTGCTTTGGCCT CCCAAAGTGCTGGGATTGCAGGTGTGAGCCACCACACCCGGCCTCTCCTT ATTTTAATGGCTCATTGTTAAACATTTACCAGCTCACTACTGCTGGGTGC AGAGGAAGAGAATGAACTAAAAAGGCAGTGAACAGACTTTCTGGAGTAAG GGGAAGTGTTACATGGATGTATAGAGTTGTAATAATCCAAGAAATTGAAC TTCAGAAACTTGTGCATTAATAGGTGAGTGCAGTGGCTCACGGCTCTAGT CCCAGCACTGCTGAGGACGAGGCAGGAGGATCGCTTGAGCCTAGGAGTGT GAGACCAGCCTGAGTGACATGGAGAAACCCTGTCTGGACAAAAAATACAA AAATTAGCCGAGTGTGGTGGCGTATGTTTGTAGCCAGGGCTACTAGGGAG GCTGAGGTGGGAGAATCGCTTGAGCCAGGGAGGTGGAGGCTGCAGAGAGT TATGATCGTGCCACTGCACTCCAGCCTGAGGCCTGGGTGACAGAGTCAGA ACTTGTCTTAAAAAGAAAAAAAAAGCCTAAAATAGGATAAAATGGGAGAA AGATTGCTAGGCAAAACAGAAGGAACATGGAAATAGCCCTGTCTCTGAAA GGGCCTGTCCTTATTTGAGGCCACATATGCATCCATCTGAATTTTGGACA AGCGGGTGGGAGCGATGAGAAGTAAAACTGAAAGGCCCAGATTGTAAAAA CCCAGGAGCAGGCTTCCCCAGGAGCAGTGTTTTGTTTGTTGTTTTGTTTT GTTTTGTTTTTTTCGAGATGGAGTCTCGGTCGGTCGCCCAGGCTGGAGTG CAATGGCGTGATCTCGGCTCACTGCAACCTCCACCTCCCGGGTTTAAGCG ATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGGCACGCATCAC CACACCCAGTTAATTTTTGTATTTTTAGTAGAGACGGGGTTTCACCATGT TGGCCAGGATAGTCTCAATCTCTTGACCTCATGATCCACCTGCCTTGGCT TCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCGCGCCCGGCCAGTTGG TTGGTTTTGTTTTTTGAGCGTGAGTCTGGCTCTGTCGCCCAGGCTGGAAT GCGATGGCACAATCTCGGCTCACTGCAACCTCCGCCTCCGGGGTTCAGTT ATCCCACCTCAGCCTCCCTAGTAGCTGGAATTACAGCCACCCGCCACCAC ACCTGTGTAATTTTTGTATTTTTAGTAGAGACGAGGTTTCACCATGTTGG CCAGGTTGGTCTCGAACTCCTGACCTCAAGTGATCAGCCCACCTCAGCTT CCCAGGGTCCTGGGATTACAGGTGTGAGCCACGGCACCTGGCAAAAAATT AAATTTTTTTTTGTTCTGTTTTATTGGAGACGGAGTCTTACTTTGTCGCC CAGGCTGGAGGGCAGTGGTGCAATCTTGGCTCACTGCAACGTCTGCCCCC CGGGTTCAAGCGATTCTCCTGCCTCAGCCGCCTGAGTAGCTGAGACTATA GGCACACACCGCCAGGCCTGGCTAATTTTTGTATTTTATTTATTTATTTG TTTGTTTGTTTGTTTGTTTGATTTTTTTGAGACGAAGTCTCGCTCTTGTC TCCCAAGCTGGAGTGCAATGGCGTGATCTTGGCTCACTGCAACCTCTGCC TCCCGGGTTCAAGCAATTCTTCTGCCTCAACCTCGCGAGTAGCTGGGATT ACAGGCACGCGCCACCATGCCCGGCTAATTTTTGTATTTTTTTGTTTTAG TAGAGACGGGGTTTCACCATGTTGGCCAGACTCGTCTTCAACTCCTGACC TCAGGTGATCCACCCGCCTCGACCTCCCAAAGTGCTGGGATTACAGGCGT GAGCCACCGCGCCCAGCCTATGACCTTTCTTATAAAGTGGTACGGCTATT GTATTAAATAGTAAGGTGGTGCTTCAAAAAGTTCAACATAGAATTACCAT ATAATCCAGTAATTCCTCTTCAGAACATATACCCAAAAGAACTCAAGGCA AGGACTCAAACAGATATTTGTACATCTGTGTTCATAGCGGCACGATTCAC CGAAGTGTCTAGTGGCAGATAAATGGATAAGCAAAATATAGTCCATGCAC ACAATAGAATATTATTCAGCCTTAAAAAGGAGGAAAATCCTGACTGGGTG CGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGATCGAGGCGGGTGG ATCACGAGGTCAGGAGATGGAGACCATCCTGGCTAACACGGTGAAACCCC GTCTCTACTAAAAATACAAAAAAATTAGCCGGGCATGGTGGTGGGCACCT GTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGG GAGGCAGAGTTTGCAGTGGGCCGAGATCGCACCACTGCACTCCAGCCTGG GTGACAGAGCGAGACTCCGTCTCAAAAAAAAAAAAAGGGAAATTTTTCTT TTTTTTTTTTTTTCTGCTCTTTTTTGGAGCAGGGCTACCCGATTGGAAGT ATGCCCGGAGTAGTCAAGTGGGTAAATTCTAACACAGGTTACAACGTGGA TCTAACACAGCTACAACAGGCACCTTGAGGACGTGGCCCTCAGTGAAATA TGCCAGCCACAAAGGGACAAAACCTGTGTGATCCTACTCATATGAAGTCC CTAGAATCATCAGATTCACAGGAAGTACGACGTTGGGTTCCAGGGGCTGG GGAGGGGGATAGGGAGTGAGGTTTCATAGGGGACAGTGTTTCAGTTTCGG AAGATGAGAAAATTCTGGAGATGGTGGTGGTGGTGGTTGCTTAATGCCGC TGAGCTGTGCATTTAGAAATGGTTAAAATGACAAGTTTTATGTTATGTGT ATTTTATAATAAAAATGTTTCAACATGCGCATAGTAATATATGCAATTTT ATTTGTCAATTAAAATAAATTTTAAAAATGTTTTAGAGTGGCCTTGTTCT GATGAAGGAGGGGGAGTAACTGACACTCTAAGGCGGTGGGGGCTAGGGGC GTGTGTCCACGCTTGCCAAAAAGATTAAATGGACTCTGGGTGGGTCTCGT CCACTGTTCTGGGGTCTTACGGGTTCTCTCAGCCCCAGCCTGGGGCACCA CAGGCTCTCAGGAGTCTGGCTACCCTGCCCACCTGTGCACGACCATCACC CCAGCCTTCATCCCTCCGTCTCCTCCCCTGCTCCCGCGCCTCATGCCCCA AGCGCTTGGGCACACGCGCCGTGGTAAGGCAAGCGAGGGCGCCCGGTCCC GGCACCGCCTCGCGTCGGTCTCCGCCCCTTTCCCCTCCGAAAGGCGGCCT TGTGCTGCTGCGCAGGCGCGGCGGCTGGGGGTGGGGTCCATCGCGGCTCC CGATCCCGTTATCGCGAGAAGCCGGTCCGCGATCTTGTGGCCGCCCCTCC CCTCCCCCTGCCTCTCGGACTCGGGCTGCGGCGTCAGCCTTCTTCGGGCC TCGGCAGCGGTAGCGGCTCGCTCGCCTCAGCCCCAGCGCCCCTCGGCTAC CCTCGGCCCAGGCCCGCAGCGCCGCCCGCCCTCGGCCGCCCCGACGCCGG CCTGGGCCGCGGCCGCAGCCCCGGGCTCGCGTAGGCGCCGACCGCTCCCG GCCCGCCCCCTATGGGCCCCGGCTAGAGGCGCCGCCGCCGCCGGCCCGCG GAGCCCCG ATG

22. CD4. CD4 (cluster of differentiation 4) is a glycoprotein found on the surface of immune cells such as T helper cells, monocytes, macrophages, and dendritic cells. In humans, the CD4 protein is encoded by the CD4 gene (Isobe et al., Proc. Natl. Acad. Sci. U.S.A. 1986; 83 (12): 4399-402). CD4+ T helper cells are white blood cells that are an essential part of the human immune system, often referred to as CD4 cells, T-helper cells or T4 cells. These helper cells send signals to other types of immune cells, including CD8 killer cells which in turn destroy and kill the infection or virus. If CD4 cells become depleted, for example in untreated HIV infection, or following immune suppression prior to a transplant, the body is left vulnerable to a wide range of infections that it would otherwise have been able to fight.

HIV-1 uses CD4 to gain entry into host T-cells and achieves this by binding to the viral envelope protein known as gp120 (Kwong et al., Nature 393 (6686): 648-59). The binding to CD4 creates a shift in the conformation of gp120 allowing HIV-1 to bind to a co-receptor expressed on the host cell. These co-receptors are chemokine receptors CCR5 or CXCR4. Following a structural change in another viral protein (gp41), HIV inserts a fusion peptide into the host cell that allows the outer membrane of the virus to fuse with the cell membrane. CD4 is also expressed in neoplasms derived from from T helper cells, e.g. peripheral T cell lymphoma and related malignant conditions and has been associated with a number of autoimmune diseases such as vitiligo and type I diabetes mellitus (Zamani et al., Clin. Exp. Dermatol. 35 (5): 521-4).

Protein: CD4 Gene: CD4 (Homo sapiens, chromosome 12, 6898638-6929976 [NCBI Reference Sequence: NC_(—)000012.11]; start site location: 6909305; strand: positive)

Gene Identification GeneID 920 HGNC 1678 HPRD 01740 MIM 186940

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 6373 GAGCCACTGCGCCCGGCCTCATTAAGGGCAT 12485 6406 CGAACAACTTCATTACAATTCGACAAGCGC 13299 6407 CGTAGTTAAGCGTGTACCAGCCCAAGGC 13189 6421 GAGCGGTGACCGTGTCTGTCTTAG 13751 6447 CGGTTTGCAGATTCCAGACCCGATGGACG 15100

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 6373 GAGCCACTGCGCCCGGCCTCATTAAGGGCAT 12485 6374 AGCCACTGCGCCCGGCCTCA 12486 6375 GCCACTGCGCCCGGCCTCAT 12487 6376 CCACTGCGCCCGGCCTCATT 12488 6377 CACTGCGCCCGGCCTCATTA 12489 6378 ACTGCGCCCGGCCTCATTAA 12490 6379 CTGCGCCCGGCCTCATTAAG 12491 6380 TGCGCCCGGCCTCATTAAGG 12492 6381 GCGCCCGGCCTCATTAAGGG 12493 6382 CGCCCGGCCTCATTAAGGGC 12494 6383 GCCCGGCCTCATTAAGGGCA 12495 6384 CCCGGCCTCATTAAGGGCAT 12496 6385 CCGGCCTCATTAAGGGCATT 12497 6386 CGGCCTCATTAAGGGCATTC 12498 6387 CGAGCCACTGCGCCCGGCCT 12484 6388 ACGAGCCACTGCGCCCGGCC 12483 6389 CACGAGCCACTGCGCCCGGC 12482 6390 GCACGAGCCACTGCGCCCGG 12481 6391 GGCACGAGCCACTGCGCCCG 12480 6392 AGGCACGAGCCACTGCGCCC 12479 6393 CAGGCACGAGCCACTGCGCC 12478 6394 ACAGGCACGAGCCACTGCGC 12477 6395 TACAGGCACGAGCCACTGCG 12476 6396 TTACAGGCACGAGCCACTGC 12475 6397 ATTACAGGCACGAGCCACTG 12474 6398 GATTACAGGCACGAGCCACT 12473 6399 GGATTACAGGCACGAGCCAC 12472 6400 GGGATTACAGGCACGAGCCA 12471 6401 TGGGATTACAGGCACGAGCC 12470 6402 CTGGGATTACAGGCACGAGC 12469 6403 GCTGGGATTACAGGCACGAG 12468 6404 TGCTGGGATTACAGGCACGA 12467 6405 GTGCTGGGATTACAGGCACG 12466 6406 CGAACAACTTCATTACAATTCGACAAGCGC 13299 6407 CGTAGTTAAGCGTGTACCAGCCCAAGGC 13189 6408 GTAGTTAAGCGTGTACCAGC 13190 6409 TAGTTAAGCGTGTACCAGCC 13191 6410 AGTTAAGCGTGTACCAGCCC 13192 6411 GTTAAGCGTGTACCAGCCCA 13193 6412 TTAAGCGTGTACCAGCCCAA 13194 6413 TAAGCGTGTACCAGCCCAAG 13195 6414 AAGCGTGTACCAGCCCAAGG 13196 6415 AGCGTGTACCAGCCCAAGGC 13197 6416 GCGTGTACCAGCCCAAGGCA 13198 6417 CGTGTACCAGCCCAAGGCAC 13199 6418 ACGTAGTTAAGCGTGTACCA 13188 6419 TACGTAGTTAAGCGTGTACC 13187 6420 GTACGTAGTTAAGCGTGTAC 13186 6421 GAGCGGTGACCGTGTCTGTCTTAG 13751 6422 AGCGGTGACCGTGTCTGTCT 13752 6423 GCGGTGACCGTGTCTGTCTT 13753 6424 CGGTGACCGTGTCTGTCTTA 13754 6425 GGTGACCGTGTCTGTCTTAG 13755 6426 GTGACCGTGTCTGTCTTAGT 13756 6427 TGACCGTGTCTGTCTTAGTT 13757 6428 GACCGTGTCTGTCTTAGTTA 13758 6429 ACCGTGTCTGTCTTAGTTAG 13759 6430 CCGTGTCTGTCTTAGTTAGC 13760 6431 CGTGTCTGTCTTAGTTAGCA 13761 6432 AGAGCGGTGACCGTGTCTGT 13750 6433 CAGAGCGGTGACCGTGTCTG 13749 6434 CCAGAGCGGTGACCGTGTCT 13748 6435 GCCAGAGCGGTGACCGTGTC 13747 6436 GGCCAGAGCGGTGACCGTGT 13746 6437 AGGCCAGAGCGGTGACCGTG 13745 6438 CAGGCCAGAGCGGTGACCGT 13744 6439 ACAGGCCAGAGCGGTGACCG 13743 6440 CACAGGCCAGAGCGGTGACC 13742 6441 TCACAGGCCAGAGCGGTGAC 13741 6442 CTCACAGGCCAGAGCGGTGA 13740 6443 GCTCACAGGCCAGAGCGGTG 13739 6444 AGCTCACAGGCCAGAGCGGT 13738 6445 TAGCTCACAGGCCAGAGCGG 13737 6446 CTAGCTCACAGGCCAGAGCG 13736 6447 CGGTTTGCAGATTCCAGACCCGATGGACG 15100 6448 CCGGTTTGCAGATTCCAGAC 15099 6449 ACCGGTTTGCAGATTCCAGA 15098 6450 CACCGGTTTGCAGATTCCAG 15097 6451 CCACCGGTTTGCAGATTCCA 15096 6452 CCCACCGGTTTGCAGATTCC 15095 6453 GCCCACCGGTTTGCAGATTC 15094 6454 GGCCCACCGGTTTGCAGATT 15093 6455 GGGCCCACCGGTTTGCAGAT 15092 6456 TGGGCCCACCGGTTTGCAGA 15091 6457 TTGGGCCCACCGGTTTGCAG 15090 6458 TTTGGGCCCACCGGTTTGCA 15089 6459 CTTTGGGCCCACCGGTTTGC 15088 6460 GCTTTGGGCCCACCGGTTTG 15087 6461 AGCTTTGGGCCCACCGGTTT 15086 6462 TAGCTTTGGGCCCACCGGTT 15085 6463 CTAGCTTTGGGCCCACCGGT 15084 6464 TCTAGCTTTGGGCCCACCGG 15083 6465 CTCTAGCTTTGGGCCCACCG 15082

Hot Zones (Relative upstream location to gene start site) 12350-12500 13100-13300 13700-13800 15000-15200

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11971) ATCTAATCTATCTATATCTGTCTATCTATCTTTATGTATCTATCTTATCT ATTGATCTATCTATCTTTTTTTTTTTTTGAGACAGAGTCACTCTGTCACC CAGGCTGGAGTGCAGTGGCACGATCTCGGCTCACTGCAACCTCCGCCTCC CGGGTTCAAGCGATTCTCCTACCTCAGCCTCCTCAGTAGCTGGGACTACC CACCACCACTCCTGGCTAATTTTTGTATTTTCAGTAGAGATAGGGTTTCA CTATGTTGGCCAGGCTGGTCTCCAACTCCTGACCTAAAGTGATCCACCCA CCTTGGTTTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCGTGCCTGG ACATATATCTATCTTTTTTTTTTTTGAGATGGAGTCTCGCTCTGTTGCCC AGGCTGGAGTGCAGTGGCGTGATTTCGGCTCACTGCAACCTCCGCCTCCC GGGTTCAAGTGATTCTCCTGCCTCAGCCTCCCAAGTAGCTGAGATTACAG ACGTGCGTCACCATGCCCAGCTAATTTTTGTATTTTTAGTAGAGATGGGA TTTCACTATGTTGGCCAGGCTGGTCTCGTACTCCCGACCTCAGGTGATCC ACTTGCCTTGGCCTCCCAAAGTGCTGGAATTACAGGTGTGAGCCACTGCA TCCGGCCTTATATATCTATCTTGTCTGTCTGACTGTCTAATCTAATTCAT CTATTTTATCTGTTTATCTTATCTATCATCTATTTATCTAATCTATCTGT CTGTATGTCTGTTTTTTTTTTGTTTTTTTTTTTTTTTTGAGATAGAGTCT TGCTCTGTCGCCGAGGCTGGAGTGCGGTGGCGCGATCTCAGCTCACTGCT GAACCTCCGCCTCCTGGGTTCTAAGCGATTCTCCTGCCTCAATCTTTGGA GTAGCTGGGATTACAGGCCCGTACCACTGTGCCCGGCTAATTTTGTATTT TTAGTAGAGAAGGGTTTCACCATGTTGGTCAGGCTTGTATTGAACTCCTG ACCTCAGGTGATCTACCCGCCTAAGCCTCCCAAAGTGCTGGGAGTACAGG TGTGAGCCACTGTGTCTGTCCCTAAATGTCTGTCTCTATCTATCTATCTA TCTATCTATCTATCTATCTATCTAATCTATCTTTCTGTCTAACCTAATCT ATTTTATCTATCTTATTCATCATCTATCTAATCTGTCTGTATGTTTATCT AATCTATTTACCTAATCTATCAATCTATCATCTAATCTATCTAATCTGTC TATCTAATCTATTTTATCTATCTATCTATCTGTCCATCCATCTATCTACC TACCTGTCTATCTCAAGCACCTACCACGTATTAAGCCCTGGCTACCTCCT CTTCCAGGCAGATGGAGTAACTGGAGGCAGCTAACAAAGATGGAGTCACT TTTCTTATCTTCTCCTAAACCACCGTAAGAGGACCAAGCCCCCACACCTT CTGAGTGCCCCATTCCTCTCCACAGATTGTGTCTTAGTGCCCAGCAGGAA ACACAGTCCACCTCCCATGGTTCAAGAGATTGTAGAAAGGGGGTTATTCA CATAGGTTAAGGGAATCAATCAATTTGAAGCACAGACACTATTAACAGCA GGAAGAGTCCTGAAGAAGTGAAAATGGTGTTTCTGGAACCCAGAGAGTGC TTGCACTCTGGATAAGGGGCCACCCCACAGAAGCTGTGGAGGGGCAGGGC TGCAGGTGAGGATGAACACACAGCTATTGACAGAAAATATGCCCAGGGCA GGGATAGAGTAGGAAAAATATCCCAGCTTCTTTCCCCCACCCTTCCATCT CATCTCTGAAAGGCACTTCCCACTGGCCAGCCCCGACTGGTGCTGGAGGG CAAGAGAGCCTATGAGCCATGTGTGGCTGTCAGCCCCTTGGTGGAGAGCC ACAGACAGGATGGAGAGTGGCTGGCAGGGCCCCGTGGGGATGAACAGCTT GGATTGGGGCGACTGGGCTTCATCCAGGCTGGGCTGGATGTGTGCATACA TTTCAGTGACCCGTTTTAGAAACAGAATTAATATGGTGAATAGAGAAAGA AGAAATCAGTGACTTTCGCTCCTCCATACAATTCAATTTGGCTTAAGTTA GCCAAAGCCATACCAAGTCCTCTCTCTATGTCTCAGCTGCTGCCAGGCTT GTGGTGGCCACACAGCTGGCTAGACTGTCATCTCTGTCCTCAAGGGGCTC AAGCTAGAGGAGGAGAGTTGAGAAACCAAATCACTATACACAAAGTAGAA GGTGGAACACACCCAGGAGCATGTCAACGGGGTGCTGTGGGACTTCAGAG TAGGCAGATCGTCACCAAGCTTCAACGGCAAAGATGCCACTGGGGGAAAG AAGGACCAAGCTTGGAAGACAGAGTAAGTCTGGAGGCAAGATCTTGTCTC ACCAGCAGGGGCCAGGTCCATGGTGACACCTTCCCCAGGCAGTCACCTCT CTGAGCCCACTTTATATCCTAGGCCTGGATTCAAAGACACTTGAGCCCTG CTCCAGCCTTCCTTTGAGGTGCTATCTTGGTGCCTTTCCTATAATCACTG CTCCAGTCCCATGTCATCTGGTCCCCAGTTACCACATCAAGCTTCCCGAA GCTCCACACAGACCATGCCACATCTTTACCAAAAAATCAGCAGTGGGTCC CCTCACCTCCAGGACAAAGCTCCAGCTCTTCGACCTGCCTGTCAATATTT GCAATCACTGCCTGCACAAATTAGCTGGGTGTTGTCATGAAAGGATCACT TGAGCCCAGGAGTTCCAGGCTGCAATGACCTATGATTGAACCACTGCACT CTGGCCTGGGTGACAGAGTGGATCTAAACTAAAAATAAAAAGATTTACAG TCAAGCCTCAAAGGCTTTTCCCATACCTTCTTCCACCATCACCTCCCTGA GCCCTCTCTTTCCTCCGAAGCCTCCTCGCACATCCCTACCACCTTTGCAC ACCTCAGAATGGGGACACCTCTCCCCTTTCCTCTCCATCTAACTTATGGT TTTCAAACTTGAGCGTGATCAGTTACCTGGAGATTTGTGAAAACCCAGAT GACTAGACCCACCCCCAGTTTCTGATTCAGCAGGTCTGGGGTGGGGCCGA GGATCTGCATTTCTAACAAGTTCCCAGGTCATGTTGCCGCTGCTACTGAT CCAGGACTTTGGGAATCGCTCCTCTAATCTACAGCTGTCCATTCCCCATG GTCCATTCAGAGCCTCTCTGCCCTGCCCCCACCACCCCCAGTCTCGCCTG TCTGCCAAGCGCACAGGAAACTCTCCTTCATCCAAACCCTGGACCAACGC CTTCTGCTTGGCCCACTCAGAGGCCTTGTAGGGTTGGTCTGATATTGGAC AGAGAAATGGCCCTCTGCTCTTTCTCCCCTGACCTCTCTGAAGGGGGCCT GCCCCTCCACACCTGTGGGTATTTCTCGCAAGGTGGAGACAAGAGACTGA GAAAAGAAATAAGACACAGAGAAAGTATAGAGGAATAAAAGTGGGCCCAG GGGACCGGCGCTCAGCAAGTGAGGACCTGCACCGGTGCTGGTCTCTGAGT TCCCTCAGTATTTATTGATCACTATCTTTACTATCTCCGCGAGGGGAATG TGGTGGGGCTATAGGGTGAAGGTGAGGAGAGGGTCAGCAGAAAAACATAT GAGCAAAGACTCTGTGTCATAAATAAGTTTAAGGAAAGGTGCTGTGCCTG GATGTGCTAGATTTATGTTTAACTTTACACAAACATCTCAGTGTAGTAAA GAGTAACAGAGCAGTATTGCCGCCATGATGTCTCGCCTCCAGACATAAGG CAGTTTTCTCCTCTCTCAAAATAGAATGTATGATCGGTTTTACACCGGGT CATTCCATTCCCAGGGACGAGCAGGAGACAGATGCCTTCCTCTTATCTCA ACCGAATAGAGGCCTTCCTCCTTCACTAATCCTCCTCAGCACAGACCCTT TACGGGTGTCGGGCTGGGGGGCTGTAAGGTCTTTCCCTTCCCATGAGGCC ATATCTCAGGCTGTCTCAGTGGGGGGAAACCTGGACAATACCTAGGCTTT CTCGGGCAGGGGTTCCTGCGGCCTTCCACAGTGTATTGTGTCTCTGGTTA ATAGAGAACGGAGAATGGTGATGACTTTCACCAAGCACACTGCCTGCAAG AACTTTTCTTTTTTTTTTTTTTTGAGACAGAGTCTTGCTCTGTCGCCCAG GCTGGAGTGCAGTGGCGCGATCTCGGCTCACTGCCACCTCTGCCTCCCGG GTTCACGCCATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGGC GCCCGCCACCACGCCCGGCTAATTTTTTTGTATTTTTTTAGTAGAGATGG GGTTTCACCGTGTTCACCAGGATGGTCTCGATCTCCTGACCTCATGATCC GCCCGCCTTGGCCTCCCAAAATGCTGGGATTACACGTGTGAGGCAAGAAC TTTTTAAAAGTGCATCTTGCGCAGCCCTAGATCCATTAAACCTTGATTCA ATACAGGACATGTTTTTGTGAGCACAGGGTTGGGACAAAAGTTACAGATT AACAGCATCTCAAAGCAGAACAATTTTTCTTAGTACAGATCAAAATGGAG TTTCTTATGTCTTCCTTTTTCTACATAGACACAGTAACAATCTGATCTCT CTTTCTTTTCCCCATACCTCTCACGCTGTATCAGGCCCCAATTCTTGGGA ACGTCACCTTAGAACTGTCCCACACATTTCTACAGCCACTTGGCTCAGGC CCTTTGCTGACCAGGATGGTTGCAGTTCTGCCTTTGGTGCCTCGCCTCCT CCAGTTCTTTCACTCAGCAGCTGCAGGGGTCCACGTGGCAAATCTAATAA TCTTCTTCTCTATAGAAAATCCTCTGCTGGCTCTCTAGTGCCCAGGATCC AGTCCCAGCATCTCAGCACGGCCTTCAAGCATTTCCACGTCCTGGCCTGG CTCCATGGTCTCCCCGCCAATTTGCCACCTTCTCCATGCATCCTTTTCTG ATCCCCTCCTCACTCATCCCAGCAAAGAACCCCCTCCTGGCCTGAGCATA GCATTTCGTGGTGTGTATCTCAGAGCATCCAGTTAGGGGTGTGCAAGTTT ACTTTGTTACTGGCTGATGTTGTGAAGTCCCAAGTTGTTGGTGCCGCAAA CAAAAAATTGGACATGACACACACAAATAGCAAAGCAGCAAAAGTTTATT AAGCACAGTACGATCCACTATGGATCAAGGATGACCTGCGAATGGTATCA GCATCACTTTGCTATATTTCATGGCCTTTTCTATGTGTTTTTTTTCTCTT TTTCCTCAAGCTGCCTAAGCTTTAGCCAGCATGTGCCTTTTGGTTGACAG GTGGGTTGCTTAGTTTCTTGGCCTCTGTGTGTTTACGTGTCATTTCCTTC CCATAGTTTTAAGTACATGCATGATATGCACTCTGTAGGCATGAACCTTA AGTAGCTAATTACTATACGGGGTCATTTTGAGGATATCTTTTCTCTGTAG TACATGTGCATCTTTTTTTGCAGTGGTGCAATCTTGGCTCACTGCAACCT CCTCCTCCCTGGTTCAAGTGATTCTCCTGCCTCAGCTTCCTAAGCACCTG AGACTACAGGTGCATGCCACCACGCCCGGCTAATTTTTGTATTTTTAGTA GAGATGGGGTTTCACCATGTTGGCCAGGCTGATCTCGAACTCCTGACCGC AAGTGATCCACCCACCTCGGCCTCCCAAAGCACTGGGATTACAGGCATGA GCCACCGCACCCAGCCTAGTATATGCCCATCTCTTAGGAGCTGCTCCTAA CTGGTTTGGTTTGGATCTAGCCAGCCATGGGGCTCCTTATTCACTTATTT ATCTTCTGTTTTTGCTCACCTGCCTCTTTCTCTTGCTTCTGCTCCTACTC ATTCCTTCCTTAATCCAACCTCCAATTCCCTCTGCTATTCTCCTGCCTCA AGTTCACTAGGCTGGCTGCAAGGGTCCTGAGGGAGAGGTTGTGTATCGCC CCTGTATACTCCAGGTCCAGTAAATGTTTGCTGACTAATGATTGGCATTT CCCTCAGGCCCTGCCATTTCTGTGGGCTCAGGTCCCTACTGGCTCAGGCC CCTGCCTCCCTCGGCAAGGCCACA ATG

23) WNT1 WNT1 (wingless-type MMTV integration site family, member 1) is a member of the WNT protein family of secreted molecules that are involved in intercellular signaling during development. WNT proteins have been shown to have regulatory roles in the cell fate process and have been associated with tumorigenesis. WNT proteins stimulate either the canonical or non-canonical intracellular signal transduction cascades. WNT proteins bind to the extracellular Frizzled (Fz) receptor family. Binding of WNT to the Fz and low density lipoprotein related protein 5/6 receptor complex, disrupts downstream protein complexes which inhibits the destruction of β-catenin. β-catenin enters the nucleus and complexes with TCF to initiate WNT-related gene expression. WNT1 has been associated multiple cancers including hepatitis B virus-related and hepatitis C virus-related hepatocellular carcinoma, gastric cancer, pancreatic cancer, breast cancer, and lung cancer.

Protein: Wnt-1 Gene: WNT1 (Homo sapiens, chromosome 12, 49372236-49376396 [NCBI Reference Sequence: NC_(—)000012.11]; start site location: 49372434; strand: positive)

Gene Identification GeneID 7471 HGNC 12774 HPRD 01276 MIM 164820

Targeted Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 6466 CGCGCGCCCGCCTCACTCAGCTGAGCG 442 6537 CGTCATTCTGTTGCCCTTTGTACCTCG 1226 6545 CGCCACGGGCGCATCCATCCCTCCTGGG 4454 6579 CACCGCCCTCTAGCCGCCTGCGGG 4960 6580 TTGCGGCGACTTTGGTTGTTGCCCGCGACGGT 34

Target Shift Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 6466 CGCGCGCCCGCCTCACTCAGCTGAGCG 442 6467 GCGCGCCCGCCTCACTCAGC 443 6468 CGCGCCCGCCTCACTCAGCT 444 6469 GCGCCCGCCTCACTCAGCTG 445 6470 CGCCCGCCTCACTCAGCTGA 446 6471 GCCCGCCTCACTCAGCTGAG 447 6472 CCCGCCTCACTCAGCTGAGC 448 6473 CCGCCTCACTCAGCTGAGCG 449 6474 CGCCTCACTCAGCTGAGCGT 450 6475 GCCTCACTCAGCTGAGCGTC 451 6476 CCTCACTCAGCTGAGCGTCC 452 6477 CTCACTCAGCTGAGCGTCCG 453 6478 TCACTCAGCTGAGCGTCCGG 454 6479 CACTCAGCTGAGCGTCCGGA 455 6480 ACTCAGCTGAGCGTCCGGAG 456 6481 CTCAGCTGAGCGTCCGGAGC 457 6482 TCAGCTGAGCGTCCGGAGCC 458 6483 CAGCTGAGCGTCCGGAGCCC 459 6484 AGCTGAGCGTCCGGAGCCCG 460 6485 GCTGAGCGTCCGGAGCCCGT 461 6486 CTGAGCGTCCGGAGCCCGTC 462 6487 TGAGCGTCCGGAGCCCGTCG 463 6488 GAGCGTCCGGAGCCCGTCGA 464 6489 AGCGTCCGGAGCCCGTCGAG 465 6490 GCGTCCGGAGCCCGTCGAGG 466 6491 CGTCCGGAGCCCGTCGAGGA 467 6492 GTCCGGAGCCCGTCGAGGAC 468 6493 TCCGGAGCCCGTCGAGGACT 469 6494 CCGGAGCCCGTCGAGGACTA 470 6495 CGGAGCCCGTCGAGGACTAG 471 6496 GGAGCCCGTCGAGGACTAGC 472 6497 GAGCCCGTCGAGGACTAGCA 473 6498 AGCCCGTCGAGGACTAGCAT 474 6499 GCCCGTCGAGGACTAGCATC 475 6500 CCCGTCGAGGACTAGCATCC 476 6501 CCGTCGAGGACTAGCATCCG 477 6502 CGTCGAGGACTAGCATCCGC 478 6503 GTCGAGGACTAGCATCCGCC 479 6504 TCGAGGACTAGCATCCGCCA 480 6505 CGAGGACTAGCATCCGCCAG 481 6506 GAGGACTAGCATCCGCCAGG 482 6507 AGGACTAGCATCCGCCAGGG 483 6508 GGACTAGCATCCGCCAGGGG 484 6509 GACTAGCATCCGCCAGGGGG 485 6510 ACTAGCATCCGCCAGGGGGC 486 6511 CTAGCATCCGCCAGGGGGCG 487 6512 TAGCATCCGCCAGGGGGCGC 488 6513 AGCATCCGCCAGGGGGCGCG 489 6514 GCATCCGCCAGGGGGCGCGG 490 6515 CATCCGCCAGGGGGCGCGGC 491 6516 ATCCGCCAGGGGGCGCGGCG 492 6517 TCCGCCAGGGGGCGCGGCGA 493 6518 CCGCCAGGGGGCGCGGCGAG 494 6519 ACGCGCGCCCGCCTCACTCA 441 6520 CACGCGCGCCCGCCTCACTC 440 6521 CCACGCGCGCCCGCCTCACT 439 6522 CCCACGCGCGCCCGCCTCAC 438 6523 TCCCACGCGCGCCCGCCTCA 437 6524 CTCCCACGCGCGCCCGCCTC 436 6525 CCTCCCACGCGCGCCCGCCT 435 6526 CCCTCCCACGCGCGCCCGCC 434 6527 ACCCTCCCACGCGCGCCCGC 433 6528 CACCCTCCCACGCGCGCCCG 432 6529 ACACCCTCCCACGCGCGCCC 431 6530 GACACCCTCCCACGCGCGCC 430 6531 GGACACCCTCCCACGCGCGC 429 6532 GGGACACCCTCCCACGCGCG 428 6533 TGGGACACCCTCCCACGCGC 427 6534 TTGGGACACCCTCCCACGCG 426 6535 CTTGGGACACCCTCCCACGC 425 6536 CCTTGGGACACCCTCCCACG 424 6537 CGTCATTCTGTTGCCCTTTGTACCTCG 1226 6538 GCGTCATTCTGTTGCCCTTT 1225 6539 TGCGTCATTCTGTTGCCCTT 1224 6540 ATGCGTCATTCTGTTGCCCT 1223 6541 TATGCGTCATTCTGTTGCCC 1222 6542 GTATGCGTCATTCTGTTGCC 1221 6543 TGTATGCGTCATTCTGTTGC 1220 6544 GTGTATGCGTCATTCTGTTG 1219 6545 CGCCACGGGCGCATCCATCCCTCCTGGG 4454 6546 GCCACGGGCGCATCCATCCC 4455 6547 CCACGGGCGCATCCATCCCT 4456 6548 CACGGGCGCATCCATCCCTC 4457 6549 ACGGGCGCATCCATCCCTCC 4458 6550 CGGGCGCATCCATCCCTCCT 4459 6551 GGGCGCATCCATCCCTCCTG 4460 6552 GGCGCATCCATCCCTCCTGG 4461 6553 GCGCATCCATCCCTCCTGGG 4462 6554 CGCATCCATCCCTCCTGGGC 4463 6555 CCGCCACGGGCGCATCCATC 4453 6556 ACCGCCACGGGCGCATCCAT 4452 6557 CACCGCCACGGGCGCATCCA 4451 6558 TCACCGCCACGGGCGCATCC 4450 6559 CTCACCGCCACGGGCGCATC 4449 6560 GCTCACCGCCACGGGCGCAT 4448 6561 AGCTCACCGCCACGGGCGCA 4447 6562 GAGCTCACCGCCACGGGCGC 4446 6563 TGAGCTCACCGCCACGGGCG 4445 6564 CTGAGCTCACCGCCACGGGC 4444 6565 GCTGAGCTCACCGCCACGGG 4443 6566 AGCTGAGCTCACCGCCACGG 4442 6567 CAGCTGAGCTCACCGCCACG 4441 6568 GCAGCTGAGCTCACCGCCAC 4440 6569 CGCAGCTGAGCTCACCGCCA 4439 6570 GCGCAGCTGAGCTCACCGCC 4438 6571 AGCGCAGCTGAGCTCACCGC 4437 6572 CAGCGCAGCTGAGCTCACCG 4436 6573 GCAGCGCAGCTGAGCTCACC 4435 6574 GGCAGCGCAGCTGAGCTCAC 4434 6575 GGGCAGCGCAGCTGAGCTCA 4433 6576 TGGGCAGCGCAGCTGAGCTC 4432 6577 GTGGGCAGCGCAGCTGAGCT 4431 6578 GGTGGGCAGCGCAGCTGAGC 4430 6579 CACCGCCCTCTAGCCGCCTGCGGG 0 6580 TTGCGGCGACTTTGGTTGTTGCCCGCGACGGT 34 6581 TGCGGCGACTTTGGTTGTTG 35 6582 GCGGCGACTTTGGTTGTTGC 36 6583 CGGCGACTTTGGTTGTTGCC 37 6584 GGCGACTTTGGTTGTTGCCC 38 6585 GCGACTTTGGTTGTTGCCCG 39 6586 CGACTTTGGTTGTTGCCCGC 40 6587 GACTTTGGTTGTTGCCCGCG 41 6588 ACTTTGGTTGTTGCCCGCGA 42 6589 CTTTGGTTGTTGCCCGCGAC 43 6590 TTTGGTTGTTGCCCGCGACG 44 6591 TTGGTTGTTGCCCGCGACGG 45 6592 TGGTTGTTGCCCGCGACGGT 46 6593 GGTTGTTGCCCGCGACGGTG 47 6594 GTTGTTGCCCGCGACGGTGG 48 6595 TTGTTGCCCGCGACGGTGGG 49 6596 TGTTGCCCGCGACGGTGGGA 50 6597 GTTGCCCGCGACGGTGGGAC 51 6598 TTGCCCGCGACGGTGGGACG 52 6599 TGCCCGCGACGGTGGGACGG 53 6600 GCCCGCGACGGTGGGACGGG 54 6601 CCCGCGACGGTGGGACGGGA 55 6602 CCGCGACGGTGGGACGGGAC 56 6603 GTTGCGGCGACTTTGGTTGT 33 6604 AGTTGCGGCGACTTTGGTTG 32 6605 CAGTTGCGGCGACTTTGGTT 31 6606 GCAGTTGCGGCGACTTTGGT 30 6607 TGCAGTTGCGGCGACTTTGG 29 6608 CTGCAGTTGCGGCGACTTTG 28 6609 GCTGCAGTTGCGGCGACTTT 27 6610 TGCTGCAGTTGCGGCGACTT 26 6611 GTGCTGCAGTTGCGGCGACT 25 6612 TGTGCTGCAGTTGCGGCGAC 24 6613 CTGTGCTGCAGTTGCGGCGA 23 6614 TCTGTGCTGCAGTTGCGGCG 22 6615 CTCTGTGCTGCAGTTGCGGC 21 6616 GCTCTGTGCTGCAGTTGCGG 20 6617 CGCTCTGTGCTGCAGTTGCG 19 6618 CCGCTCTGTGCTGCAGTTGC 18 6619 CCCGCTCTGTGCTGCAGTTG 17 6620 GCCCGCTCTGTGCTGCAGTT 16 6621 TGCCCGCTCTGTGCTGCAGT 15 6622 TTGCCCGCTCTGTGCTGCAG 14 6623 TTTGCCCGCTCTGTGCTGCA 13 6624 CTTTGCCCGCTCTGTGCTGC 12 6625 GCTTTGCCCGCTCTGTGCTG 11 6626 GGCTTTGCCCGCTCTGTGCT 10 6627 TGGCTTTGCCCGCTCTGTGC 9 6628 CTGGCTTTGCCCGCTCTGTG 8 6629 CCTGGCTTTGCCCGCTCTGT 7 6630 GCCTGGCTTTGCCCGCTCTG 6 6631 TGCCTGGCTTTGCCCGCTCT 5 6632 CTGCCTGGCTTTGCCCGCTC 4 6633 CCTGCCTGGCTTTGCCCGCT 3 6634 GCCTGCCTGGCTTTGCCCGC 2 6635 GGCCTGCCTGGCTTTGCCCG 1

Hot Zones (Relative upstream location to gene start site)   1-1000 1050-1450 1600-1900 3300-3800 4250-4700 4750-5000

Examples

In FIG. 42, In MCF7 (human mammary breast cell line), WNT1_(—)1, WNT1_(—)2, WNT1_(—)3 produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated control values. The WNT1 sequences WNT1_(—)1, WNT1_(—)2, and WNT1_(—)3 fit the independent and dependent DNAi motif claims.

The secondary structures for WNT1_(—)1, WNT1_(—)2, and WNT1_(—)3 are shown in FIG. 43, FIG. 44, and FIG. 45.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11972) CCCGGGGAACCCAAATTATAGGCCCAGGAGGGATGGATGCGCCCGTGGCG GTGAGCTCAGCTGCGCTGCCCACCCTCCGCTTAATGCGCCTTCTGCTGCA GCACCGTAGGCCACCACCTGGAGGCACCAAAGGGTCTGCGGGCCGACTGC ATACTGGACTCTCAGGAAGGCCCCACTTTCAGCAGTCCACTCCAACAAAT CCATGGGATTACCTTAGACAGAATTTTGCCCCCTTCTGTACTCAGGCCTA ATGGATGGGCTGTGCCTTCCCAGCCCAAGGGGGCAGTGCTGCCTGCGGGT GCTTCAAAGGAGGGTAGGCTCCTCTGCCCACAAACTCTAAACCCTGGAGC CCTGCTTCCTCCCCAGATCCCAAAGTCAAGGCAAAGCCCCTCTCCCCTCT AACATCTCACCTCTAACCCTATTCCAGGGGGGTGGTTTGCTACTGATTTT CAACTTCAAGCCTTTAAAGTCATCCACGGTCAAAACTGATACAGAGAAAA ATGAAGCAGGGTAAAGGAGATTAGTAGTGGGATTCTATTTTATAAAGGGG GAGGGAAAACAAACTGAAGGAACAAATACATGGAGAGATCTGAGGAAGAG CATTTTAGAAGACAGAAAAGCAGGTGCACAGACCCTTAGAAAGGAGCATG CTTGGTTCAAAGGATTAGAAAAGAGGCCAGTGAGGCTAGTGGGGAGAAAT TCGCAAGGAAGAGAGTGGTAGGCAATGAAGCTGGAGGGCTAGGAAGGAGG CCCCTTTACTTTGAGTGACATGGGGTCTCGCTGGAAAATTTTGAGCAGAG AAATGAACTAATCAGACTTCTGTTTTAGGAAAGATGGCTCTGGGCTGGGC GCAGTGGCTCATGCCTGTAATCCCAGTACTTTGGGAGGCCGACGTGGGCA GATCACGAGGTCAGGAGTTCGAGACCAGCCTGGCCAACATGGTGAAACCC CACCTCTACTAAAAATACAAAAATTAGGTGGGAGTGTTGGTGGGCGCCTG TAATCCCAGCTACTCGGTACGCTGAGGCAGGAGAATCGCTTGAAACCGGA AGGTGGAGGTTACAGTGAGCCGAGATCATACCACTGCACTCCAGTCTGCA CAACAAGAGCAAAACTCCGTCTCAAAAAAAAAAAAAAAGAAAGAAAGAAA GAAAGAAATTGGCTCTAGTAATTAAATCAACCCTTTTGATTTTTGCAGTA AAATGGAGCACTGAAATGGAGCAATCTTGGGCAGTAATGTGGGGTCTAAT GAAGTTTTTGGGTTTTTCAGATGGGTACTATTGCAGCATGTCTGCATGCT TATGTGTATGTTCCAGGAGAGAGATAAGTGGATGATGCAGGAAAGAAAGA GGGGACAGTTGATATCATGATTATTTGATCTAAATAGAAAGTTGGGTGCT TGTTTTGGCAGCACATATACTAAAATTGGAATGATATAGAGATTAGCATG GCCCCTGCACAAGGATGACATGCAAATTTGTGAAGCATTTCATATTTTGA AAAAGAAATGTCAGCCAGGTCATAAACAGTGTGACCCAGATCTAGGGGCC TTACCCTCTTGCCCCCTACTCCTGGTGTGTGGAATGTTGGAACAAAGCAC AGTGGCTCCTTTCCTCTCTTCCCACCTCTGCTTGACAACAGTCGTCAAAG ACAGGGCTTCCATATTTTCCAGCCAGCCTCCCACCCTCACGGTGTTGTAT CAATCCACCAGGCCAAAAGATGTGACCCAGGCCCCAGTGGGAAGAAACTC ATAAGGGATAAAGGACAGGCTCCCCGTGATACATTGTCCATTTACTTGAG CTATCTATGCTGGGTGCTCTCTGCAGGGACTACTGGCTTTTGGATCTACG GAGGGTGCTGGACCACTACACCTTTTCCCTCTGGGGTGGATCCTTGGAAG GGCCAGATATACTAGGCTGGGCAAAAGGGAAGAAAAAAGGGAAAGAAGGA CATTTCTTTCTAAAATAACTTCCATCAGGCTTCATTTGGGTTAATATGCA TCTCATTTAAAACACAAGTGCCCGGGAATATTAATGAAACTTACCTGGCA TTTATTCCTTAGAGTGATTTCCCTGCCTTAGAAGGGAATCCTAGTCATTT CTGGGACTTGAGACTTTAGGTTCAGGCCTGGGGAAATTTCTCAGTCAGAA GGCATCCTAAAAGACAAGGGAGATGAAAAAAATGAGAGCTAGAACTCAAA AGGGAGGCAGAAAGGCCCAAAAAATTATTTTTACCCATCAATTTTGAGAA GGGTTCCCAGCCTGTAATTGCTGCACACTGGCAAGCAGCTGGTAAGGTCG AAAGAGCATGGGCTTTGAGTCAAATTGGTCTGGGTTTTAATCTGGCTCTA CCATTGATTCATTTATTAGACGTGGACCTTGGACAAGTGCCTGATCTATT TTTAATTCTGCAAAATGGGGAGAGAGAAGAGATCTTCCCTCCTTCCAGGG GCCATGTGTGTGGTGGTGGGGCATGATAACCAGGCTGGCAGTGCCCCCTA TTCCCCATATAGGGAAAAGCAGCCACTTTTTTTTTTTTTTTCAGATGAAG TCTTGCTCTGTAGCCCAGACTGGAGTGCAATGGCGTGATCTCGGCTCACT GCAATCTCCACCTCCCGGGTTCAAGCCATTCTCCTGCCTCAGCCTCCTGA GTAGCTGGGACTACAGGTGTGCGCCACCACACTCAGCTAATTTTTGTATT TTTATTAGAGATGGGATTCCACTATGTTGGCCAGGATGGTCTCGATCTCC TGACCTCATGATCCACCTGCCTCGGCCTCCCAAAGTGCGGGGATTATAGG CATGAGCCACCGCGCCCAGCCTGTCACTTCTTCAATAGGAGGCCTAAATG GCCTTGAAGCTGAGTAGGAGTCCCTGGGAGAGAAGAGAAAAGTGTACAAT GGATGAGATGGTCACAGGCACTCTGGGTATCCCAGTGTGGTGGGAACTAG AGCTTTAGGGAAAGACAGAAACTTGGCAGAAACATCCAAAGAGAAGCAAA CACATGGAGGCACAAGTTTCCTCATCTAGGTTCAATGTAGCCAGCAACCC TTGTCTTCCCAGTCCTCTCCATCACCATACATACAGTGGACATCCGCACC ATTTCCCATCCTTTCTGAGCCTAGGCCTCAGAGACTTAGCCACTCCAGGC TGGGTTCACCTCAATACCATCTTGGTTGTAGGCTCGGCTCTCTCCCCCAA TGACATGCACTGGTTGACACATACCACAGTGTGACACGCCATAGGATGCC ACGAGGTACAAAGGGCAACAGAATGACGCATACACACATATTTAATCTTC CCATGCACATGCTCATCCACCCACTCCACACACAGTCCAGACACTCTGCA TCCCTCAATCATGCTTCTGAGTCTCCTGTCGACAGTTGCCACCTCCTTCC TGACACACTGCCCCAGGCGGTGACTGTGACAAGGTGACTCCATGACCTTT TCTGACTTGAGCTAAATTCCAAAATTCTTTGGAAAGTTTCCTAACATCCT TCGTCAGAACAAGGAGTTTCTGCACGTACCAACACACAGGAGGATGCACC CTCAGAACACAGCACATTCTCACTCCCACCCATATTCACGTTGTTCCACT TCACACACACACACACACACACACACACACACAGCCACTTGTGCGCTTCT TCTGGCGCACATGAGCAAACTGCCTGTTGCTTTAGGTTTCTCTCCACCGC TAGGCTCCTTTTGGTTAGCTCACCCCCACAACTCATCCCCGGGATTTCCC TGACCACAGCCGCACTCACGCCCCCGTCTCCCCTTTTTCCTTCTCTGTCC AGCCATCGGGGGTTCCTGGGCGGTTAAGCATCTCCCCGGAGTCGCTGCCC AGAACCACAGCTTTCCTTCCGACACTCAGGATGGGGGAGAGAGGGGACGT CGGAGGGGCCCGGGGTGACGTCGAGGGGACAACCCCACCGCGGGCGGCGA GGCGGGCTGGGCCCCTGGCGGGCTCTCCCCGCAGCACACTCTCGCCGCGC CCCCTGGCGGATGCTAGTCCTCGACGGGCTCCGGACGCTCAGCTGAGTGA GGCGGGCGCGCGTGGGAGGGTGTCCCAAGGGGAGGGGTCCGCGGCCAGTG CAGGCCCGGAGGCGGGGGCCACCGGGCAGGGGGCGGGGGTGAGCCCCGAC GGCCAACCCGTCAGCTCTCGGCTCAGACGGGCGGGAACCACAGCCCCGCT CGCTGCCCATTGTCTGCGCCCCTAACCGGTGCGCCCTGGTGCCACAGTGC GGCCCGGAGGGGCAGCCTCCTCCCGTCACTTCAGCCAGCGCCGCAACTAT AAGAGGCGGTGCCGCCCGCCGTGGCCGCCTCAGCCCACCAGCCGGGACCG CGAGCCATGCTGTCCGCCGCCCGCCCCCAGGGTTGTTAAAGCCAGACTGC GAACTCTCGCCACTGCCGCCACCGCCGCGTCCCGTCCCACCGTCGCGGGC AACAACCAAAGTCGCCGCAACTGCAGCACAGAGCGGGCAAAGCCAGGCAG GCC ATG

24) Clusterin. Clusterin is a heterodimeric glycoprotein produced by a wide array of tissues and found in most biologic fluids. A number of physiologic functions have been proposed for clusterin based on its distribution and in vitro properties. These include complement regulation, lipid transport, sperm maturation, initiation of apoptosis, endocrine secretion, membrane protection, and promotion of cell interactions. A prominent and defining feature of clusterin is its induction in such disease states as glomerulonephritis, polycystic kidney disease, renal tubular injury, neurodegenerative conditions including Alzheimer's disease, atherosclerosis, and myocardial infarction (reviewed by Rosenberg and Silkensen, Int. J. Biochem Cell Biol. 1995: 27 (7) 633-645. Genome-wide association studies found a statistical association between a SNP within the clusterin gene and the risk of having Alzheimer's disease (Lambert et al., 2009: Nat. Genet. 41 (10): 1094-1099). Other studies, Alzheimer's patients have more clusterin in their blood (Schrijvers et al. 2011 JAMA 305 (13): 1322-1326).

Clusterin acts as cell-survival protein and is over-expressed in response to anti-cancer agents. An antisense approach to inhibiting clusterin (Curtisen) has shown promising results in combination with currently available chemotherapies in several tumor types. The FDA granted Custirsen two Fast Track Designations as a treatment in combination with first-line and second-line docetaxel for progressive metastatic prostate cancer.

Protein: Clusterin Gene: CLU (Homo sapiens, chromosome 8, 27454434-27472328 [NCBI Reference Sequence: NC_(—)000008.10]; start site location: 27468088; strand: negative)

Gene Identification GeneID 1191 HGNC 2095 HPRD — MIM 185430

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 6636 CGTCCCGCCCACCTGCTGCCTGCAGCAG 78 6660 CGACAATCAGCGAGGCACACAGGCT 330 6689 CGGAGAGTAGAGAGGGTTCGCAGTGGCCC 718 6690 CCACGGGGCACAGGCCATAGCCCCG 890 6709 CTCGTGCTCTCAGGCGGCGGTTGCGCCG 3865 6752 CCGGGAGGTGGGGGCCGGTGCAGCACCGG 4260 6753 TCGCGTGCCCATCTGGGAGCCCCTCTCACG 4395

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 6636 CGTCCCGCCCACCTGCTGCCTGCAGCAG 78 6637 GTCCCGCCCACCTGCTGCCT 79 6638 TCCCGCCCACCTGCTGCCTG 80 6639 CCCGCCCACCTGCTGCCTGC 81 6640 CCGCCCACCTGCTGCCTGCA 82 6641 CGCCCACCTGCTGCCTGCAG 83 6642 GCGTCCCGCCCACCTGCTGC 77 6643 GGCGTCCCGCCCACCTGCTG 76 6644 TGGCGTCCCGCCCACCTGCT 75 6645 CTGGCGTCCCGCCCACCTGC 74 6646 GCTGGCGTCCCGCCCACCTG 73 6647 TGCTGGCGTCCCGCCCACCT 72 6648 CTGCTGGCGTCCCGCCCACC 71 6649 CCTGCTGGCGTCCCGCCCAC 70 6650 GCCTGCTGGCGTCCCGCCCA 69 6651 AGCCTGCTGGCGTCCCGCCC 68 6652 CAGCCTGCTGGCGTCCCGCC 67 6653 ACAGCCTGCTGGCGTCCCGC 66 6654 GACAGCCTGCTGGCGTCCCG 65 6655 AGACAGCCTGCTGGCGTCCC 64 6656 TAGACAGCCTGCTGGCGTCC 63 6657 CTAGACAGCCTGCTGGCGTC 62 6658 GCTAGACAGCCTGCTGGCGT 61 6659 AGCTAGACAGCCTGCTGGCG 60 6660 CGACAATCAGCGAGGCACACAGGCT 330 6661 GACAATCAGCGAGGCACACA 331 6662 ACAATCAGCGAGGCACACAG 332 6663 CAATCAGCGAGGCACACAGG 333 6664 AATCAGCGAGGCACACAGGC 334 6665 ATCAGCGAGGCACACAGGCT 335 6666 TCAGCGAGGCACACAGGCTT 336 6667 CAGCGAGGCACACAGGCTTT 337 6668 AGCGAGGCACACAGGCTTTC 338 6669 GCGAGGCACACAGGCTTTCT 339 6670 CGAGGCACACAGGCTTTCTG 340 6671 CCGACAATCAGCGAGGCACA 329 6672 CCCGACAATCAGCGAGGCAC 328 6673 CCCCGACAATCAGCGAGGCA 327 6674 TCCCCGACAATCAGCGAGGC 326 6675 CTCCCCGACAATCAGCGAGG 325 6676 CCTCCCCGACAATCAGCGAG 324 6677 TCCTCCCCGACAATCAGCGA 323 6678 ATCCTCCCCGACAATCAGCG 322 6679 CATCCTCCCCGACAATCAGC 321 6680 ACATCCTCCCCGACAATCAG 320 6681 CACATCCTCCCCGACAATCA 319 6682 CCACATCCTCCCCGACAATC 318 6683 GCCACATCCTCCCCGACAAT 317 6684 AGCCACATCCTCCCCGACAA 316 6685 AAGCCACATCCTCCCCGACA 315 6686 CAAGCCACATCCTCCCCGAC 314 6687 CCAAGCCACATCCTCCCCGA 313 6688 TCCAAGCCACATCCTCCCCG 312 6689 CGGAGAGTAGAGAGGGTTCGCAGTGGCCC 718 6690 CCACGGGGCACAGGCCATAGCCCCG 890 6691 CACGGGGCACAGGCCATAGC 891 6692 ACGGGGCACAGGCCATAGCC 892 6693 CGGGGCACAGGCCATAGCCC 893 6694 GCCACGGGGCACAGGCCATA 889 6695 AGCCACGGGGCACAGGCCAT 888 6696 GAGCCACGGGGCACAGGCCA 887 6697 TGAGCCACGGGGCACAGGCC 886 6698 CTGAGCCACGGGGCACAGGC 885 6699 CCTGAGCCACGGGGCACAGG 884 6700 CCCTGAGCCACGGGGCACAG 883 6701 GCCCTGAGCCACGGGGCACA 882 6702 TGCCCTGAGCCACGGGGCAC 881 6703 CTGCCCTGAGCCACGGGGCA 880 6704 GCTGCCCTGAGCCACGGGGC 879 6705 GGCTGCCCTGAGCCACGGGG 878 6706 TGGCTGCCCTGAGCCACGGG 877 6707 CTGGCTGCCCTGAGCCACGG 876 6708 GCTGGCTGCCCTGAGCCACG 875 6709 CTCGTGCTCTCAGGCGGCGGTTGCGCCG 3865 6710 TCGTGCTCTCAGGCGGCGGT 3866 6711 CGTGCTCTCAGGCGGCGGTT 3867 6712 GTGCTCTCAGGCGGCGGTTG 3868 6713 TGCTCTCAGGCGGCGGTTGC 3869 6714 GCTCTCAGGCGGCGGTTGCG 3870 6715 CTCTCAGGCGGCGGTTGCGC 3871 6716 TCTCAGGCGGCGGTTGCGCC 3872 6717 CTCAGGCGGCGGTTGCGCCG 3873 6718 TCAGGCGGCGGTTGCGCCGG 3874 6719 CAGGCGGCGGTTGCGCCGGG 3875 6720 AGGCGGCGGTTGCGCCGGGG 3876 6721 GGCGGCGGTTGCGCCGGGGC 3877 6722 GCGGCGGTTGCGCCGGGGCC 3878 6723 CGGCGGTTGCGCCGGGGCCC 3879 6724 GGCGGTTGCGCCGGGGCCCC 3880 6725 GCGGTTGCGCCGGGGCCCCT 3881 6726 CGGTTGCGCCGGGGCCCCTG 3882 6727 GGTTGCGCCGGGGCCCCTGG 3883 6728 GTTGCGCCGGGGCCCCTGGC 3884 6729 TTGCGCCGGGGCCCCTGGCT 3885 6730 TGCGCCGGGGCCCCTGGCTC 3886 6731 GCGCCGGGGCCCCTGGCTCA 3887 6732 CGCCGGGGCCCCTGGCTCAG 3888 6733 GCCGGGGCCCCTGGCTCAGC 3889 6734 CCGGGGCCCCTGGCTCAGCT 3890 6735 CGGGGCCCCTGGCTCAGCTG 3891 6736 GCTCGTGCTCTCAGGCGGCG 3864 6737 AGCTCGTGCTCTCAGGCGGC 3863 6738 GAGCTCGTGCTCTCAGGCGG 3862 6739 GGAGCTCGTGCTCTCAGGCG 3861 6740 TGGAGCTCGTGCTCTCAGGC 3860 6741 TTGGAGCTCGTGCTCTCAGG 3859 6742 GTTGGAGCTCGTGCTCTCAG 3858 6743 GGTTGGAGCTCGTGCTCTCA 3857 6744 TGGTTGGAGCTCGTGCTCTC 3856 6745 GTGGTTGGAGCTCGTGCTCT 3855 6746 TGTGGTTGGAGCTCGTGCTC 3854 6747 TTGTGGTTGGAGCTCGTGCT 3853 6748 ATTGTGGTTGGAGCTCGTGC 3852 6749 AATTGTGGTTGGAGCTCGTG 3851 6750 GAATTGTGGTTGGAGCTCGT 3850 6751 AGAATTGTGGTTGGAGCTCG 3849 6752 CCGGGAGGTGGGGGCCGGTGCAGCACCGG 4260 6753 TCGCGTGCCCATCTGGGAGCCCCTCTCACG 4395 6754 CGCGTGCCCATCTGGGAGCC 4396 6755 GCGTGCCCATCTGGGAGCCC 4397 6756 CGTGCCCATCTGGGAGCCCC 4398 6757 CTCGCGTGCCCATCTGGGAG 4394 6758 ACTCGCGTGCCCATCTGGGA 4393 6759 AACTCGCGTGCCCATCTGGG 4392 6760 GAACTCGCGTGCCCATCTGG 4391 6761 TGAACTCGCGTGCCCATCTG 4390 6762 CTGAACTCGCGTGCCCATCT 4389 6763 CCTGAACTCGCGTGCCCATC 4388 6764 GCCTGAACTCGCGTGCCCAT 4387 6765 AGCCTGAACTCGCGTGCCCA 4386 6766 GAGCCTGAACTCGCGTGCCC 4385 6767 AGAGCCTGAACTCGCGTGCC 4384 6768 AAGAGCCTGAACTCGCGTGC 4383 6769 GAAGAGCCTGAACTCGCGTG 4382 6770 GGAAGAGCCTGAACTCGCGT 4381 6771 GGGAAGAGCCTGAACTCGCG 4380 6772 AGGGAAGAGCCTGAACTCGC 4379 6773 TAGGGAAGAGCCTGAACTCG 4378

Hot Zones (Relative upstream location to gene start site)  1-950 1000-1300 2050-3000 3550-4500

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11973) AATGTGAAGGTTAAGGTCAGTAGGGCCAGGGAACTGTGAGATTGTGTCTT GGACTGGGACAGACAGCCGGGCTAACCGCGTGAGAGGGGCTCCCAGATGG GCACGCGAGTTCAGGCTCTTCCCTACTGGAAGCGCCGAGCGGCCGCACCT CAGGGTCTCTCCTGGAGCCAGCACAGCTATTCGTGGTGATGATGCGCCCC CCGGCGCCCCCAGCCCGGTGCTGCACCGGCCCCCACCTCCCGGCTTCCAG AAAGCTCCCCTTGCTTTCCGCGGCATTCTTTGGGCGTGAGTCATGCAGGT TTGCAGCCAGCCCCAAAGGGGGTGTGTGCGCGAGCAGAGCGCTATAAATA CGGCGCCTCCCAGTGCCCACAACGCGGCGTCGCCAGGAGGAGCGCGCGGG CACAGGGTGCCGCTGACCGGTGAGATGTCCCCGTCTTCCCTACCCTTGAG CAGAGCCACACCAGGACGGATGGGCGGGCAGGGGATGGCAGCCAGGCAGA GAGGGATGACACAGCTCGCAGTCACAACCCCTGCGCTTTCGACGGAGCCC AGGAAGCCAGGGAGGGGAGGTGGCCGGAGCCCCATCACCAGGCAGCTGAG CCAGGGGCCCCGGCGCAACCGCCGCCTGAGAGCACGAGCTCCAACCACAA TTCTGTGGTGGGGGGGTAAATAGAACAGATATAATGATCATCCTTTCGCA AAGATGGGGAAACTGAGACCTGGAGACCTGCCGCGTTGCGGGAGACCCAG GCTAGCAGGTGACAGAGCTGGCCTGCACCGAGCTCCTTCCTGCAGCATAT CCTCTGCGAAGATGCGGATCTCTCAGTTGTGGCTTTCGGCTTGCATGCAT GAGTCATCTAGTTTTCTTCTAAATTCTCTAGCTCTCTGGACACTGTTGCC TGTAAGTATGAGGCTGCGGATTTCAGTATATGCTGCAACCACCGAAATCC GACTTTTTCTGCCTCCTAATGCATCTGAGGTGCATCAGAGAAAAGTCACA CAAGATCCACCAGGCCTCAGACCTCTGATTCCACAGTCTCATTTTACAGA TGATAATCTGAGGCCTGGAGAGGTTTAGGACTGGTGCCAACACTAAACAG CAAATAAGTATCAGAATTGGGATTCGAGCCAAAGCCTCTTGACCTTCCAG AATTTCTGGACCTAGTTAAAAAAAATATGATTTTTATTATTATTTTTTAA ACGGAGAGGTTAGGAATTTAAAGGAAAGTACAGATACTATATAAAAAAAG ATGCCCATGAAAATGTTAAGTTATAATAATAGTGGAGCATTGGGCACAAC TGAAATGGCCAATCTTGTGAGAATGGTAAAATAAACTTAGGTCCGTGAGT AAGTGGAGTATTACATAGCCATAAAAGTATGCCCTTAAAGAATATTTGAA GATGGTGAATGTGAAGAATCTTGTATAAACTGCATGGAAGACAGAAGGAA ATATACCACAGTGCTAACCTTTGCCTCTGGGTGATATGAATTACCGGTGA TTATTTTTCTTATTTTCCTTTTGGTTTAGTTTTCTCCATTTGAAGAAGCA GATAGGAGCCGGGGCTTTGGGATTGAAACCCTCACCATCTGTGTGCCCTC TTCACTGTCTTCCCATCCTCCCCACGGCTCCCTGTTCACAGTCATTGATT TTCTTTCTTTCTTTTCTCTCTTTTTTTTTTTTTTTCCTGAGACCAAGTCT CACTCTGTTGCCCAGGCTGGAGTAGAGTAGCGCCATCTCGGCTCACTGCA ACCTCCGCCATCCGGGTTCAAGCAGTTCTCATGCCTCAGCCTCTGAGTAG CTGGGACTACAGGCGCATGCTGCTACATCCGGCTAATTTTTGTATTTTTA GTAGAGACATGGTTTCACCACCTTGGCCAGGCTGGTCTCGAACTCCTGAT CTCAAGTAATCCGCCTGTCTTGGCCTCCCAAAGTGCTGGGGTGACAGGTG TGAATCAATGCGCCCTGCCAGGTCATTGATTTTCTTAAGCCTCCAGCCCT GCCCTGCTTGGAAACGTTTTGGGAAGCTGCTCAGTTCAAAGTTCCCAGGA GGGTGTGCCTGGAGGGGAGTTGCTCCCAAAGTCTGCCTGCTCCCCCCGCC CCCCCTGCCCCCCACCCCCCGCCATCTTCTCCTCCTCCTCTTCCCCTGAG CAGCCCCTTTGTCCACAGAACCGGCCTTTTCTGGTAGAAGGAGCAAGGCC AAGTGGTTTAAGCCTTCTTAGGGAGAATGAGGCTGTGTGGTAGTGCTGGG GACTCGAGGGCCTTGGCCTTGGCATGGCTCTTCCACCCAGGGCAGCTGGC AGCCAGGCTCCCAGGAGGCAGAGGAGATGAGGGGGGAGGTGAGTCCGAGC AAAGGAAAGGAGGTCGGCTGTGCAGTCACGGTTCTAGAACATTCCTTGGA TCAGCAGCATCCATATCACCTGCAGACTGGCTGGAAAAGCAGTCTCAGAA CCAACATTATAACCAGCCCTGCAGTGATTCATAAGTACTTTAAAAAGTGG TCAATCATTTCAGCAAAGCAGAGCCACACAGTCCGGGGGACCACAGGTGG CCTCTGTGTGCTTGTCTCGGTTTTCCTGCCCCTCTCCAGACATGTTGATT AGACACTGCCAATGCCCAGCCTCAGACCTCAGTCTAATTTGGAAGTAGTC AGAATTTACTATGATTACATAAGACCCTCGTGTTTACAGAACACATTCCC CTCTCTGAGGTCTGGATTAGATCCATTTTACAGATGAAGAAACTGAGGCT CAGATATTTAAGTGACTTGGAATCAAGGAAAGAATACTGGACTAGGGGTC GGGAGGGCTGGGCTCTCATCCCAGGGTTACCATGAGCATGCTGTGGACTC TAGGGAGTCCCATGCCCTCTCTGGGCTTCAGCCTCACCGCTAGGGTAGAG AGGTTGGGTGAGAGAACGACCTCCTTCCCAGGTCTGAGCTGGATGGTTCA CCAGGGACCCCAGGCTCCCTGGAGCAGACTCTGTGCCCGCTGCTGAGTCT GGAATTCCTTTCCTGTATCTTGCCTTTGGCTGCCCCATTCTTCATGGCCC AGCACCCTGTCTTCTGGTCAGAACCTAGTTCTGAATGGGTTTTTCCAGAA GTTGTTGCTTTCAGGGGCCCCTGGCAGAGAGGTGTTTCTGGCTGGCTTTG TCTCTCTGGCATGACAAAGGCTCTGTTCCTGCTGGAGGCATTTCAGGGCT CAGTGGGCAGCTGGGGCAGAGCCCGTGAGACCACAGCCTTCCTGGTGAGC CCGGTCTCCGCCCCCTACCCCATCTCTGGGGAAGGCGCTGACCCCATCTC TTCTCCCACGCTGCTCCCTGGCTCTTTGCGCCTGATTACTTCTCATGAGA GGCACTCCTTGTTAATGTGCTACTGAGTGTCCAGATGGGCCTGCTGGGCT GAGCGGGCTTTGGATGTGAACCATTTCAGGAAGGGGAACCCCATCGTCCT GTTGGTTCTGTGATGGCAAATGGGTGAGCTCAGATAAGCAGTTCTTGGGA GGGGCATGGTGGGGGTGGAGTGCAGGGGGAGGGGTTTCTGTTTTATGCAA CAGCCTCAGCTTCTGGGAAAGGGTCCATTGTGTAAGACCGGGGCTATGGC CTGTGCCCCGTGGCTCAGGGCAGCCAGCCCAGTGGTGGCAGGAACACTGG CAGGGCAGCCTGCTGTCGGCTTAGAGGGGATGGGCAGTGTGGAGGGCCTG GCAGAGCAAGAGGACTCATCCTTCCAAAGGGACTTTCTCTGGGAAGCCTG CTCCTCGGGCCACTGCGAACCCTCTCTACTCTCCGAAGGGAATTGTCCTT CCTGGCTTCCACTACTTCCACCCCTGAATGCACAGGCAGCCCGGCCCAAG TCTCCCACTAGGGATGCAGATGGATTCGGTGTGAAGGGCTGGCTGCTGTT GCCTCCGGCTCTTGAAAGTCAAGTTCAGGTGGTGCTGAGACTCCCTGGGG GCTGCAGCGCTGTGGTGAATGGGGAGCGTCTGCTGGGGTGAAGGTTTAGG TGCACATTGCAGAGGACGTGGCTGGTCTCTGGGATGCAGTCCCTCTGTGG AGGTGGCATGGGGAGGGACGGATGCATGACCTAAGGGGGGTATTTTCAGT GTCTGACATGATCGATACCACTCTGGACAAGGAGGCCAGGATGCAGAAAG CCTGTGTGCCTCGCTGATTGTCGGGGAGGATGTGGCTTGGACAAGAGCCT GGTTCCTCCGATGCCAGGGTTCTTGTTTCTTCCACTCAACATTGCTGTCC TGCAGTCCCTCCCTCCCTGCACCTCCTGCCTTCGCTTTCATTCGAGGTGT CCATGGCAAGTCTGGTCATTTCCCCCCATTTCCTCAGGAATAAAAGTGCA GCAGTGCCTGCTGTGGGGACAGCTGAGGGCAGTGAGGCCCTGGGGAGCTG CTGCAGGCAGCAGGTGGGCGGGACGCCAGCAGGCTGTCTAGCTGTTCCCA TGATGGTCTCCTGTTCTCTGCAGAGGCGTGCAAAGACTCCAGAATTGGAG GC ATG

25. NRAS. The neuroblastoma RAS viral oncogene homolog (N-ras) oncogene is a member of the Ras gene family. It is mapped on chromosome 1, and it is activated in HL60, a promyelocytic leukemia line. The mammalian ras gene family consists of the harvey and kirsten ras genes (HRAS and KRAS), an inactive pseudogene of each (c-Hras2 and c-Kras1) and the N-ras gene. They differ significantly only in the C-terminal 40 amino acids. These ras genes have GTP/GDP binding and GTPase activity, and their normal function may be as G-like regulatory proteins involved in the normal control of cell growth. Mutations which change amino acid residues 12, 13 or 61 activate the potential of N-ras to transform cultured cells and are implicated in a variety of human tumors. The N-ras gene specifies two main transcripts of 2 Kb and 4.3 Kb. The difference between the two transcripts is a simple extension through the termination site of the 2 Kb transcript. The N-ras gene consists of seven exons (-I, I, II, III, IV, V, VI). The smaller 2 Kb transcript contains the VIa exon, and the larger 4.3 Kb transcript contains the VIb exon which is just a longer form of the VIa exon. Both transcripts encode identical proteins as they differ only the 3′ untranslated region (reviewed in Marshall et al., 1982 Nature 299 (5879): 171-3 and Shimizu et al., 1983 PNAS 80 (2): 383-7).

Protein: NRAS Gene: NRAS (Homo sapiens, chromosome 1, 115247085-115259515 [NCBI Reference Sequence: NC_(—)000001.10]; start site location: 115258781; strand: negative)

Gene Identification GeneID 4893 HGNC 7989 HPRD 01273 MIM 164790

Targeted Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 6774 CCCCGCCCTCAGCCTAAGCAATGGA 234 6793 GACCCCGGAACCGCCATGAACAGCCC 559 6818 CCCGCTACGTAATCAGTCGGCGCCCCA 613 6961 AACGCAAAAACACCGGATTAATATCGGCCT 142 6963 ATAAACGGCCTCTTTACCCAGAGATCA 850 6971 CGCCACCTTAAGTTTTTCCAGGCTGC 1779

Target Shift Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 6774 CCCCGCCCTCAGCCTAAGCAATGGA 234 6775 CCCGCCCTCAGCCTAAGCAA 235 6776 CCGCCCTCAGCCTAAGCAAT 236 6777 CGCCCTCAGCCTAAGCAATG 237 6778 GCCCCGCCCTCAGCCTAAGC 233 6779 GGCCCCGCCCTCAGCCTAAG 232 6780 GGGCCCCGCCCTCAGCCTAA 231 6781 TGGGCCCCGCCCTCAGCCTA 230 6782 TTGGGCCCCGCCCTCAGCCT 229 6783 CTTGGGCCCCGCCCTCAGCC 228 6784 CCTTGGGCCCCGCCCTCAGC 227 6785 TCCTTGGGCCCCGCCCTCAG 226 6786 GTCCTTGGGCCCCGCCCTCA 225 6787 AGTCCTTGGGCCCCGCCCTC 224 6788 CAGTCCTTGGGCCCCGCCCT 223 6789 ACAGTCCTTGGGCCCCGCCC 222 6790 AACAGTCCTTGGGCCCCGCC 221 6791 CAACAGTCCTTGGGCCCCGC 220 6792 TCAACAGTCCTTGGGCCCCG 219 6793 GACCCCGGAACCGCCATGAACAGCCC 559 6794 ACCCCGGAACCGCCATGAAC 560 6795 CCCCGGAACCGCCATGAACA 561 6796 CCCGGAACCGCCATGAACAG 562 6797 CCGGAACCGCCATGAACAGC 563 6798 CGGAACCGCCATGAACAGCC 564 6799 GGAACCGCCATGAACAGCCC 565 6800 GAACCGCCATGAACAGCCCC 566 6801 AACCGCCATGAACAGCCCCC 567 6802 ACCGCCATGAACAGCCCCCA 568 6803 CCGCCATGAACAGCCCCCAC 569 6804 CGCCATGAACAGCCCCCACC 570 6805 AGACCCCGGAACCGCCATGA 558 6806 GAGACCCCGGAACCGCCATG 557 6807 GGAGACCCCGGAACCGCCAT 556 6808 TGGAGACCCCGGAACCGCCA 555 6809 TTGGAGACCCCGGAACCGCC 554 6810 GTTGGAGACCCCGGAACCGC 553 6811 TGTTGGAGACCCCGGAACCG 552 6812 ATGTTGGAGACCCCGGAACC 551 6813 AATGTTGGAGACCCCGGAAC 550 6814 AAATGTTGGAGACCCCGGAA 549 6815 AAAATGTTGGAGACCCCGGA 548 6816 AAAAATGTTGGAGACCCCGG 547 6817 GAAAAATGTTGGAGACCCCG 546 6818 CCCGCTACGTAATCAGTCGGCGCCCCA 613 6819 CCGCTACGTAATCAGTCGGC 614 6820 CGCTACGTAATCAGTCGGCG 615 6821 GCTACGTAATCAGTCGGCGC 616 6822 CTACGTAATCAGTCGGCGCC 617 6823 TACGTAATCAGTCGGCGCCC 618 6824 ACGTAATCAGTCGGCGCCCC 619 6825 CGTAATCAGTCGGCGCCCCA 620 6826 GTAATCAGTCGGCGCCCCAG 621 6827 TAATCAGTCGGCGCCCCAGG 622 6828 AATCAGTCGGCGCCCCAGGC 623 6829 ATCAGTCGGCGCCCCAGGCG 624 6830 TCAGTCGGCGCCCCAGGCGC 625 6831 CAGTCGGCGCCCCAGGCGCC 626 6832 AGTCGGCGCCCCAGGCGCCT 627 6833 GTCGGCGCCCCAGGCGCCTG 628 6834 TCGGCGCCCCAGGCGCCTGA 629 6835 CGGCGCCCCAGGCGCCTGAG 630 6836 GGCGCCCCAGGCGCCTGAGT 631 6837 GCGCCCCAGGCGCCTGAGTC 632 6838 CGCCCCAGGCGCCTGAGTCC 633 6839 GCCCCAGGCGCCTGAGTCCC 634 6840 CCCCAGGCGCCTGAGTCCCC 635 6841 CCCAGGCGCCTGAGTCCCCG 636 6842 CCAGGCGCCTGAGTCCCCGC 637 6843 CAGGCGCCTGAGTCCCCGCC 638 6844 AGGCGCCTGAGTCCCCGCCC 639 6845 GGCGCCTGAGTCCCCGCCCC 640 6846 GCGCCTGAGTCCCCGCCCCG 641 6847 CGCCTGAGTCCCCGCCCCGG 642 6848 GCCTGAGTCCCCGCCCCGGC 643 6849 CCTGAGTCCCCGCCCCGGCC 644 6850 CTGAGTCCCCGCCCCGGCCA 645 6851 TGAGTCCCCGCCCCGGCCAC 646 6852 GAGTCCCCGCCCCGGCCACG 647 6853 AGTCCCCGCCCCGGCCACGT 648 6854 GTCCCCGCCCCGGCCACGTG 649 6855 TCCCCGCCCCGGCCACGTGG 650 6856 CCCCGCCCCGGCCACGTGGG 651 6857 CCCGCCCCGGCCACGTGGGC 652 6858 CCGCCCCGGCCACGTGGGCC 653 6859 CGCCCCGGCCACGTGGGCCT 654 6860 GCCCCGGCCACGTGGGCCTC 655 6861 CCCCGGCCACGTGGGCCTCC 656 6862 CCCGGCCACGTGGGCCTCCG 657 6863 CCGGCCACGTGGGCCTCCGA 658 6864 CGGCCACGTGGGCCTCCGAA 659 6865 GGCCACGTGGGCCTCCGAAC 660 6866 GCCACGTGGGCCTCCGAACC 661 6867 CCACGTGGGCCTCCGAACCA 662 6868 CACGTGGGCCTCCGAACCAC 663 6869 ACGTGGGCCTCCGAACCACG 664 6870 CGTGGGCCTCCGAACCACGA 665 6871 GTGGGCCTCCGAACCACGAG 666 6872 TGGGCCTCCGAACCACGAGT 667 6873 GGGCCTCCGAACCACGAGTC 668 6874 GGCCTCCGAACCACGAGTCA 669 6875 GCCTCCGAACCACGAGTCAT 670 6876 CCTCCGAACCACGAGTCATG 671 6877 CTCCGAACCACGAGTCATGC 672 6878 TCCGAACCACGAGTCATGCG 673 6879 CCGAACCACGAGTCATGCGG 674 6880 CGAACCACGAGTCATGCGGC 675 6881 GAACCACGAGTCATGCGGCA 676 6882 AACCACGAGTCATGCGGCAG 677 6883 ACCACGAGTCATGCGGCAGG 678 6884 CCACGAGTCATGCGGCAGGC 679 6885 CACGAGTCATGCGGCAGGCC 680 6886 ACGAGTCATGCGGCAGGCCG 681 6887 CGAGTCATGCGGCAGGCCGC 682 6888 GAGTCATGCGGCAGGCCGCA 683 6889 AGTCATGCGGCAGGCCGCAC 684 6890 GTCATGCGGCAGGCCGCACC 685 6891 TCATGCGGCAGGCCGCACCC 686 6892 CATGCGGCAGGCCGCACCCA 687 6893 ATGCGGCAGGCCGCACCCAG 688 6894 TGCGGCAGGCCGCACCCAGA 689 6895 GCGGCAGGCCGCACCCAGAC 690 6896 CGGCAGGCCGCACCCAGACC 691 6897 GGCAGGCCGCACCCAGACCC 692 6898 GCAGGCCGCACCCAGACCCG 693 6899 CAGGCCGCACCCAGACCCGC 694 6900 AGGCCGCACCCAGACCCGCC 695 6901 GGCCGCACCCAGACCCGCCC 696 6902 GCCGCACCCAGACCCGCCCC 697 6903 CCGCACCCAGACCCGCCCCT 698 6904 CGCACCCAGACCCGCCCCTC 699 6905 GCACCCAGACCCGCCCCTCC 700 6906 CACCCAGACCCGCCCCTCCC 701 6907 ACCCAGACCCGCCCCTCCCA 702 6908 CCCAGACCCGCCCCTCCCAC 703 6909 CCAGACCCGCCCCTCCCACA 704 6910 CAGACCCGCCCCTCCCACAC 705 6911 AGACCCGCCCCTCCCACACG 706 6912 GACCCGCCCCTCCCACACGG 707 6913 ACCCGCCCCTCCCACACGGG 708 6914 CCCGCCCCTCCCACACGGGA 709 6915 CCGCCCCTCCCACACGGGAC 710 6916 CGCCCCTCCCACACGGGACG 711 6917 GCCCCTCCCACACGGGACGT 712 6918 CCCCTCCCACACGGGACGTT 713 6919 CCCTCCCACACGGGACGTTT 714 6920 CCTCCCACACGGGACGTTTC 715 6921 CTCCCACACGGGACGTTTCA 716 6922 TCCCACACGGGACGTTTCAA 717 6923 CCCACACGGGACGTTTCAAT 718 6924 CCACACGGGACGTTTCAATA 719 6925 CACACGGGACGTTTCAATAA 720 6926 GCCCGCTACGTAATCAGTCG 612 6927 CGCCCGCTACGTAATCAGTC 611 6928 CCGCCCGCTACGTAATCAGT 610 6929 CCCGCCCGCTACGTAATCAG 609 6930 CCCCGCCCGCTACGTAATCA 608 6931 GCCCCGCCCGCTACGTAATC 607 6932 GGCCCCGCCCGCTACGTAAT 606 6933 CGGCCCCGCCCGCTACGTAA 605 6934 CCGGCCCCGCCCGCTACGTA 604 6935 TCCGGCCCCGCCCGCTACGT 603 6936 TTCCGGCCCCGCCCGCTACG 602 6937 CTTCCGGCCCCGCCCGCTAC 601 6938 ACTTCCGGCCCCGCCCGCTA 600 6939 CACTTCCGGCCCCGCCCGCT 599 6940 GCACTTCCGGCCCCGCCCGC 598 6941 GGCACTTCCGGCCCCGCCCG 597 6942 CGGCACTTCCGGCCCCGCCC 596 6943 GCGGCACTTCCGGCCCCGCC 595 6944 AGCGGCACTTCCGGCCCCGC 594 6945 GAGCGGCACTTCCGGCCCCG 593 6946 GGAGCGGCACTTCCGGCCCC 592 6947 AGGAGCGGCACTTCCGGCCC 591 6948 AAGGAGCGGCACTTCCGGCC 590 6949 CAAGGAGCGGCACTTCCGGC 589 6950 CCAAGGAGCGGCACTTCCGG 588 6951 ACCAAGGAGCGGCACTTCCG 587 6952 CACCAAGGAGCGGCACTTCC 586 6953 CCACCAAGGAGCGGCACTTC 585 6954 CCCACCAAGGAGCGGCACTT 584 6955 CCCCACCAAGGAGCGGCACT 583 6956 CCCCCACCAAGGAGCGGCAC 582 6957 GCCCCCACCAAGGAGCGGCA 581 6958 AGCCCCCACCAAGGAGCGGC 580 6959 CAGCCCCCACCAAGGAGCGG 579 6960 ACAGCCCCCACCAAGGAGCG 578 6961 AACGCAAAAACACCGGATTAATATCGGCCT 142 6962 GAACGCAAAAACACCGGATT 141 6963 ATAAACGGCCTCTTTACCCAGAGATCA 850 6964 TAAACGGCCTCTTTACCCAG 851 6965 AAACGGCCTCTTTACCCAGA 852 6966 AACGGCCTCTTTACCCAGAG 853 6967 ACGGCCTCTTTACCCAGAGA 854 6968 CGGCCTCTTTACCCAGAGAT 855 6969 GATAAACGGCCTCTTTACCC 849 6970 AGATAAACGGCCTCTTTACC 848 6971 CGCCACCTTAAGTTTTTCCAGGCTGC 1779 6972 GCGCCACCTTAAGTTTTTCC 1778 6973 GGCGCCACCTTAAGTTTTTC 1777 6974 AGGCGCCACCTTAAGTTTTT 1776 6975 AAGGCGCCACCTTAAGTTTT 1775 6976 TAAGGCGCCACCTTAAGTTT 1774 6977 ATAAGGCGCCACCTTAAGTT 1773 6978 TATAAGGCGCCACCTTAAGT 1772 6979 CTATAAGGCGCCACCTTAAG 1771 6980 ACTATAAGGCGCCACCTTAA 1770 6981 TACTATAAGGCGCCACCTTA 1769 6982 ATACTATAAGGCGCCACCTT 1768 6983 GATACTATAAGGCGCCACCT 1767 6984 TGATACTATAAGGCGCCACC 1766 6985 TTGATACTATAAGGCGCCAC 1765

Hot Zones (Relative upstream location to gene start site)  1-950 1700-2000

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11974) CCACATCCACAAAGCACACCATTAATCCACTATGATCAAGTTGGGGGGAA TCTGGTGAAGGGTTCTGAATATCTCCCTCTTCATCCCTCCCGAAATCTGG AATACTTATTCTATTGAGCTATTACACCAGTTTTAACACCTTCCTCGTGT TATGTTTAAAAAAATAAATAAATTTAAGAAAACCATTTTAAATAATGCAC AGTTGCAGCCTGGAAAAACTTAAGGTGGCGCCTTATAGTATCAATTTTAG GAGCTTTATTTGGTGCATTTAACGCAACTGGTAATTGCAGAATCCACTTT GCCTGTGTAAGTGAAAAATATAGACTGTTATCTTGTTGGCCCTATGAAAT TCTGCACTTTTCATTATATACTCTACCTTCATTAATTACTTCTGGCAAGA TGTTCTGCCTTAGCACTCAGTTGCATTCTTTTCCTTTTTCTTCCTGTTCA TTATGCTTTAATTCTGAGGACCATATGAGGGTAGAATATATTATCTTTTA AAAATTACAAAAATTTGTATAGGCAAACCATTTCTTAAAGTTGATGGCCA AATTTTAAAATGTTATTTTTCATATCATTTATAATCTTGTCACAATCCAC TTAAAGAAGTTTGGTTATATTTCAGTGAAAATTTTCTTCCAGAGTAGGTT TTTTTTCGTGGGTTGGGGGGTAACTTTACTACAATTAGTAAGTATGGTGC AGAATTTCATGCAAATGAGGAGTGCCAGCAGTGTGATAATTTAAACATAT TTAAACAAAAACAAAAAAAATGAATGCACAAACTTGCTGCTGCTTAGATC ACTGCAGCTTCTAGGACCCGGTTTCTTTTACTGATTTAAAAACAAAACAA AAAAAAATAAAAAAGTTGTGCCTGAAATGAATCTTGTTTTTTTTTATAAG TAGCCGCCTGGTTACTGTGTCCTGTAAAATACAGACACTTGACCCTTGGT GTAGCTTCTGTTCAACTTTATATCACGGGAATGGATGGGTCTGATTTCTT GGCCCTCTTCTTGAATTGGCCATATACAGGGTCCCTGGCCAGTGGACTGA AGGCTTTGTCTAAGATGACAAGGGTCAGCTCAGGGGATGTGGGGGAGGGC GGTTTTATCTTCCCCCTTGTCGTTTGAGGTTTTGATCTCTGGGTAAAGAG GCCGTTTATCTTTGTAAACACGAAACATTTTTGCTTTCTCCAGTTTTCTG TTAATGGCGAAAGAATGGAAGCGAATAAAGTTTTACTGATTTTTGAGACA CTAGCACCTAGCGCTTTCATTATTGAAACGTCCCGTGTGGGAGGGGCGGG TCTGGGTGCGGCCTGCCGCATGACTCGTGGTTCGGAGGCCCACGTGGCCG GGGCGGGGACTCAGGCGCCTGGGGCGCCGACTGATTACGTAGCGGGCGGG GCCGGAAGTGCCGCTCCTTGGTGGGGGCTGTTCATGGCGGTTCCGGGGTC TCCAACATTTTTCCCGGCTGTGGTCCTAAATCTGTCCAAAGCAGAGGCAG TGGAGCTTGAGGTAAGTTTATCTCATGCATAGTGTTCGGCTTTGGGCTGT GGAATGTTCAGGCGTTTCACTGATGCCAGAAATGGAGCAGAATCTATCAG CTGGAGACAAAGGCCTTGGGCGGGGGTCCTTCCATTTGGTGCCTACGTGG GGAGATCTTTGGAGACAGAAGGGAGAATGGGAAGGAGTTGCGGCCTGGAG GCTTCCTGCTAGAGCTGAGAAGCCTTCGGGGAGTAATAGGAAGGGGGATT TCCATTGCTTAGGCTGAGGGCGGGGCCCAAGGACTGTTGAAAAATAGCTA AGGATGGGGGTTGCTAGAAAACTACTCCAGAAGTGTGAGGCCGATATTAA TCCGGTGTTTTTGCGTTCTCTAGTCACTTTAAGAACCAAATGGAAGGTCA CACTAGGGTTTTCATTTCCATTGATTATAGAAAGCTTTAAAGTACTGTAG ATGTGGCTCGCCAATTAACCCTGATTACTGGTTTCCAACAGGTTCTTGCT GGTGTGAAATG

26. EZH2. Histone-lysine N-methyltransferase (EZH2) is an enzyme that belongs to the Polycomb-group (PcG) family. PcG family members form multimeric protein complexes, which are involved in maintaining the transcriptional repressive state of genes over successive cell generations. EZH2 acts mainly as a gene silencer; it performs this role by the addition of three methyl groups to Lysine 27 of histone 3, a modification leading to chromatin condensation (Cao et al., 2002, Science 298 (5595): 1039-43). Mutations in in the EZH2 gene cause Weaver syndrome (Gibson et al., 2011: Am J Hum Genet 90 (1): 110-8). EZH2 overproduction may cause cancer due to increase in histone methylation. This histone methylation may play a role in silencing the expression of tumor suppressor genes, which may cause certain cancers. The microRNA produced by miR-101 normally inhibits translation of the messenger RNA coding for EZH2. Loss of this microRNA gene therefore leads to increased production of EZH2.

Protein: EZH2 Gene: EZH2 (Homo sapiens, chromosome 7, 148504464-148581441 [NCBI Reference Sequence: NC_(—)000007.13]; start site location: 148544390; strand: negative)

Gene Identification GeneID 2146 HGNC 3527 HPRD 03342 MIM 601573

Targeted Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 6986 TCCCGACAAGGGGTGACAGAGGC 1002 7002 CGTGAATTCAAGAGTTGCTTAGGCC 1059 7003 GACTACCGGTGCCCGCCACCACGCCAGGC 2856 7035 CCCCCGCCAACCCCACAGCGGATGCCCCC 34593459 CGCCAACCCCACAGCGGATGC

Target Shift Sequences Relative upstream location to Sequence gene No: Sequence (5′-3′) start site 6986 TCCCGACAAGGGGTGACAGAGGC 1002 6987 ATCCCGACAAGGGGTGACAG 1001 6988 CATCCCGACAAGGGGTGACA 1000 6989 GCATCCCGACAAGGGGTGAC 999 6990 AGCATCCCGACAAGGGGTGA 998 6991 CAGCATCCCGACAAGGGGTG 997 6992 ACAGCATCCCGACAAGGGGT 996 6993 CACAGCATCCCGACAAGGGG 995 6994 GCACAGCATCCCGACAAGGG 994 6995 AGCACAGCATCCCGACAAGG 993 6996 CAGCACAGCATCCCGACAAG 992 6997 GCAGCACAGCATCCCGACAA 991 6998 TGCAGCACAGCATCCCGACA 990 6999 CTGCAGCACAGCATCCCGAC 989 7000 GCTGCAGCACAGCATCCCGA 988 7001 TGCTGCAGCACAGCATCCCG 987 7002 CGTGAATTCAAGAGTTGCTTAGGCC 1059 7003 GACTACCGGTGCCCGCCACCACGCCAGGC 2856 7004 ACTACCGGTGCCCGCCACCA 2857 7005 CTACCGGTGCCCGCCACCAC 2858 7006 TACCGGTGCCCGCCACCACG 2859 7007 ACCGGTGCCCGCCACCACGC 2860 7008 CCGGTGCCCGCCACCACGCC 2861 7009 CGGTGCCCGCCACCACGCCA 2862 7010 GGTGCCCGCCACCACGCCAG 2863 7011 GTGCCCGCCACCACGCCAGG 2864 7012 TGCCCGCCACCACGCCAGGC 2865 7013 GCCCGCCACCACGCCAGGCT 2866 7014 CCCGCCACCACGCCAGGCTA 2867 7015 CCGCCACCACGCCAGGCTAA 2868 7016 CGCCACCACGCCAGGCTAAT 2869 7017 GCCACCACGCCAGGCTAATT 2870 7018 CCACCACGCCAGGCTAATTT 2871 7019 CACCACGCCAGGCTAATTTT 2872 7020 ACCACGCCAGGCTAATTTTT 2873 7021 CCACGCCAGGCTAATTTTTT 2874 7022 CACGCCAGGCTAATTTTTTG 2875 7023 GGACTACCGGTGCCCGCCAC 2855 7024 GGGACTACCGGTGCCCGCCA 2854 7025 TGGGACTACCGGTGCCCGCC 2853 7026 GTGGGACTACCGGTGCCCGC 2852 7027 GGTGGGACTACCGGTGCCCG 2851 7028 AGGTGGGACTACCGGTGCCC 2850 7029 TAGGTGGGACTACCGGTGCC 2849 7030 GTAGGTGGGACTACCGGTGC 2848 7031 AGTAGGTGGGACTACCGGTG 2847 7032 AAGTAGGTGGGACTACCGGT 2846 7033 CAAGTAGGTGGGACTACCGG 2845 7034 CCAAGTAGGTGGGACTACCG 2844 7035 GACCGCCCCCCGCCAACCCCACAGCGG 3453 7036 ACCGCCCCCCGCCAACCCCA 3454 7037 CCGCCCCCCGCCAACCCCAC 3455 7038 CGCCCCCCGCCAACCCCACA 3456 7039 GCCCCCCGCCAACCCCACAG 3457 7040 CCCCCCGCCAACCCCACAGC 3458 7041 CCCCCGCCAACCCCACAGCG 3459 7042 CCCCGCCAACCCCACAGCGG 3460 7043 CCCGCCAACCCCACAGCGGA 3461 7044 CCGCCAACCCCACAGCGGAT 3462 7045 CGCCAACCCCACAGCGGATG 3463 7046 GCCAACCCCACAGCGGATGC 3464 7047 CCAACCCCACAGCGGATGCC 3465 7048 CAACCCCACAGCGGATGCCT 3466 7049 AACCCCACAGCGGATGCCTA 3467 7050 ACCCCACAGCGGATGCCTAA 3468 7051 CCCCACAGCGGATGCCTAAA 3469 7052 CCCACAGCGGATGCCTAAAG 3470 7053 CCACAGCGGATGCCTAAAGC 3471 7054 CACAGCGGATGCCTAAAGCT 3472 7055 ACAGCGGATGCCTAAAGCTG 3473 7056 CAGCGGATGCCTAAAGCTGC 3474 7057 AGCGGATGCCTAAAGCTGCA 3475 7058 GCGGATGCCTAAAGCTGCAG 3476 7059 CGGATGCCTAAAGCTGCAGA 3477 7060 AGACCGCCCCCCGCCAACCC 3452 7061 AAGACCGCCCCCCGCCAACC 3451 7062 CAAGACCGCCCCCCGCCAAC 3450 7063 CCAAGACCGCCCCCCGCCAA 3449 7064 CCCAAGACCGCCCCCCGCCA 3448 7065 TCCCAAGACCGCCCCCCGCC 3447 7066 CTCCCAAGACCGCCCCCCGC 3446 7067 TCTCCCAAGACCGCCCCCCG 3445 7068 ATCTCCCAAGACCGCCCCCC 3444 7069 TATCTCCCAAGACCGCCCCC 3443 7070 TTATCTCCCAAGACCGCCCC 3442 7071 CTTATCTCCCAAGACCGCCC 3441 7072 ACTTATCTCCCAAGACCGCC 3440 7073 CACTTATCTCCCAAGACCGC 3439 7074 CCACTTATCTCCCAAGACCG 3438

Hot Zones (Relative upstream location to gene start site)  1-300  900-1100 2600-3100 3400-4200

Examples

In FIG. 46, In MCF7 (human mammary breast cell line), EZH2_(—)2 (271) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The EZH2 sequence EZH2_(—)2 (271) fits the independent and dependent DNAi motif claims.

The secondary structure for EZH2_(—)2 (271) is shown in FIG. 47.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11975) CACAGGCTCAAGAGATCCTCCCACCTCAGCTTCCTGAGTAGTTGGGACCA CAGGTGTGCGCCACTACACCTGGCTTGCTTGCTTGCTTATTTATTGATTT GAGATGGGAGTCTCACTATATTGCCTAGGCTGGTCTTGAACTCCTGGGCT CAATCCTCCAACCTTGGCCTCCCAAAATGCTGGGTTTACAGGCTTGAACC ACTGTACGTGGCCTTGAATCTGTGTTTTAATACTATGCTTACTTGGCTGT GGTGTTGTGAAAAGATCACTGAAAATGGAGTCAGAGGCCTGATTTGAGCC AGTCGTTTGTTGTGGGGGAAGGAGGTCAGGGGAGCTAACATCTAAAGGCT CACTATATGCCAGGCACAGAACCAAGTGTGTTTGCATGTATATTTCGTTT TTGTTGCCAGACTTTGAGGTAGGTTTTATGGATAAGGTCTTTAAGGCAAT ATCAGCTTCCTTTTAAAAAAGAAATTCCGGAAACTGAGTTTTAGGCTGAA GATCTCTAACTGGTAGTAGGGACAACTGAACCACAGGGTCCTAACTGACC CTGCGATTTATCTCCTTTTGCGGGGGGTTTCTTGATAATAGGGTGCACTT TACCTCATTTTTTGGCTCAAGCATGGATAGGCCACCCTTCCTTTTCATAC CTATAGCTAAGCTTTACAAATGATATGCTGATAAGATACAAGCTACTCGT TATTCATGTGGGTTAATAGACCTGTTTGTTTGCTTGTTTTTAAGTCTATA GCCGCCCCACCCCCAATCTACAATTTCACCTTCTAAGGTTTTAGTTACTC ATTCAAACTGCAGTCTGAAAATGTTACGATATTTTGAGAGAGAGAAGACT CTAGCTACGTAACTTTTGTAACAATATATTGTTATAATTGTTCATTTTAC CATTAGTTATTGCTGTCAGTCTCTTACTGTGCCTTATTTATAAATTAAAC TTCATGGGTATGTACGTATAGGAAAAAACATGGTATATTTAGAGTTTAGT ACTATCTGCAGCTTTAGGCATCCGCTGTGGGGTTGGCGGGGGGCGGTCTT GGGAGATAAGTGGGGACTACTGTACAATTATCAGGCACACACAGGCTCTG GGATTTTACAAATGAGTAAAAGTGGTTCTTGCTGTTGAAGCACTTACAGT GGGAATAGAGTGAAATACATGAAAATGTGATTTTAATATGTTATAAATGC TATGATGGTGGGAGTTTGTTTTGTGTAAAACATCCTTTTAATTGGTACTT TAAATTTTAATATTCTTTCACAGGTCTACCTATTTAGTCTTACACTTTCA AAGAACTACCTGGATGCTGTAGATTTTCATGATATACTTTATTAGGTATG TTATTAATGGTAGAAACAGCATGGAAAGTCTTCCAGAATATTAGACAAGG ACAGTTCTAGTACTAAAACATAAAATGCTAACTAATGTCTTCATCAAGAC ATAAAATATGTATCTTAAAAAATAAATTGTAAGCCAGGCGCAGTGGCTCA CACCTGTAATCCCAGCACTTTGGGAGGCTGAGGCGGGTGGATCACAAGGT CAGGAGATTGAGACCATCCTGGCTAACACGGTGAAACCCTGTCTCTACTA AAAATACAAAAAATTAGCCTGGCGTGGTGGCGGGCACCGGTAGTCCCACC TACTTGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCAGAGCT TGCAGTGAGCGGAGATTGCACCACCGCACTCCAGCTTGGGGGACAGAGTG AGACTCCATCTCAAAAAAAAAGAAATTGTAATAACACCCACATTATACAT CAGTGAAAACTAAACACGTTACTACCCTAGGCCTTATTGCACAGGGGTGC TACCTCCAAGGAGAAATTTGTCTAGGCAGCAGATGGACTAGAGGTGATTA GCCTATGAGCGAATGAGGCTACAGATCATTCCTTTTTATCTGATTCCTTT TCTTTCTAGTTCCTAGGCCTTGGAAGCACTAAGTGGTCTTAAGTAATTTG CATAGAATTAGTTGAGTTCATCTGTTAACTAACTAGCAGATAGGAAGAAA ACTATTGTCATGAAATTATTTAAAAAATAATAATGCTCCAGTTTCTTCTC ATCTTTGATGTCCTTTGGTCCTACCTCACTGCCTTCCTAACACCATTTTC TGCTTTACCTCAAAGCTGGGGTCATCTTGAGTTTAGCCTGCTTAATCCGA GTGACTGTCAGCTTTATTCCACTTTAGCAACTCGCAGGCAAGGCCACACT TGGAAACTTTTCACTTGGAATAGTTCTATCTTGGTGATTTCATCAGCCTT CTTTATGTCAAATACACTCAAATTCCTGCCCATTCTTATCTCTTCGTTCC CTTCAGGTCCTTAGTCCTTTTAATTTGTGACTTTCATTCTCCAGGTCCAT TCTTATTAAATGTCTGCCAGCCAGACTTTAGTTGCCCCCTGTCCAGCTTT CTCTTGCTCAGACCTAAGATTTCTTTAGGTTCTTTCTTTGCCTTTTGAAA TCCAGCTCAGCTTTTAAGATTGAGTTCCTTGTTACCTTTCCTGCCATCCT TCTGCAGTTCCTAATATTCTTTTCTTTCTCCCAAAGTGCTTTTGTATAAA CAGTCAGCCTTCCATATCCGTGAGTTCCAAATCCATGGATCCTGAATTCA TGGATTTAACCAGCTGCAGATAAAAAATATTCAGAAAAAAAAAGATGGTT GCATCTGTACTGAACATGTCTGTACTGTTTTGCTTGTCATTATTTTCTAA ACAATACAGTATAACAACTATTTACATGGCATTTACATTGTATTAGGGAT TATAAGTAATCTAGAGGTGATTTAAAGTATATGGGAGTCTCTTATATCCC AGGAAGCCAGGTAAAAAAAAAAAGTATATGGGAGGCTATGCATAGGTGAT ATGCAAATATTACACCACTTTATATCACGGACTTTTGAGCATCTGTGGAT TTTGGTATCCGAGGGGTGTCCTGGAACCAGTTCCCCAGGGATACTGAAGT ATGTCTGTCTATCTCATACTATATTTTTCCCTTTGTCTTAGGTAGACTAT CAGCTCCATAGGGGCAAGGATTTAATAATATTTGTATATTCATTTTATTC AAGTTCGTATACACTGCTTGGGTCATAATATTTATTACATGCTTGAGAAA ATGAATTTCTTCGCCCCTTTGTTACAGCTCTGAGTAAACAGCCATCTGCC TTCTCTGTCATCTGTTGGTGGTTGAGTATTTCTGTAGAAAGTTACCCATT GGCCTCAGGACTCTTACTCTAAATCTTCTTCTTAGGCAGTTTTCTCTGTG CATGAAGTTTTTATGTAAACAAATAGATGAAGCCTGCCCTACTCATTTAT TTGCTCAAGCCAGAAAGTCACCTTCTTCTTCACTTTCCATATTTAAATCA TCATTTGGTGGAATTTTGGCCTAAGCAACTCTTGAATTCACGTACTTTTC CCTGTCATCGCCAGTGTGGTGTAGAAGCCTCTGTCACCCCTTGTCGGGAT GCTGTGCTGCAGCATCATCTAACCTGGTTGCAGTTATTCTTTCACTCCCT CACCGCACACCCTTTTACTTAAAACACTAAAAGTGGCTTCTCATTGTTCT TAAGATAAAGCACAAATTGTTAGTGTGGCCTGTAAAGCTTTGCATAGCCT GACAGAGAATGTCCTGCTAATAATTTGAAGGTACAGGATGATTTTAATAC TTTAGGAGAAAATGTTCTAGGAAAAGACGCTTGTTTAGACTTAAGGTGAG GACTCTGCAGTATGAATTAGACATCTGGTGAACTATAAGCTGTCCCCGCA TTTAAACATAATTGGTTCTGAGAGCCTGCAACTAAAGATAAGGCAGAAGA ATTTACTTTGCATTTCCTGCATTCCTCTTTTCGCTTGATAGCAGAAACCC CTCATGTTAATAAAGGTGGCACAAGAGGCAAAAATACAGACTTTATCACA GTGTTTAAGGAGAGGTGCATGATTAAGTGTGTGGGGAGAGAGTACCTTTG TACATTTTATTATATGGTGAACTGTATGTTTTCTACTTTTAGTACTGTTT GTAAATTTTACTTCTTCTTGGATTTACCTTTTTCAGTTATATTATTCCAT TATGCCTTGCTACTGTAACAGCTAATGATGAAAAACAGGATCTGTCTTTA TATTTTCTTCCCTCCACAAATGTGGATCTCATAGAGTTGAAAACTAGGTT GTGATATAGTATAGTATACCTAATTCCTGTAATGGGATCATGTTCCTATA ATATGGCCGCAATTTAGTGTAGAATTTTTGTAAATAAAAGTGTATTTTAA GTTTAACTTAAACTTTCAATGAAGTGTTTTAAGGATTTAACCATGCAGCA CAAATGAGCACCTTTCTGTAAATGCCAACAGTGTAATATGTGTCATTTCT TCACTGATTGTTAGTTTGCTGCGGATTAAAACACAGGTGATCATATTCAG GCTGGTTAGATTAGTGATTTTAATATGAAACCATTGCTTTTAGAATAATC ATG

27. HDACs, such as HDAC1.

Histone deacetylases (HDACs) are part of a vast family of enzymes that have crucial roles in numerous biological processes, largely through their repressive influence on transcription (reviewed by Haberland et al, 2009 Nature Reviews Genetics 10, 32-42. HDAC1 is an enzyme that belongs the histone deacetylase family and is a component of the histone deacetylase complex (Taunton et al, Science 272 (5260): 408-11) Histone acetylation and deacetylation, is catalyzed by multisubunit complexes and is key in the expression of gene expression. It also interacts with retinoblastoma tumor-suppressor protein and this complex is a key element in the control of cell proliferation and differentiation. Together with metastasis-associated protein-2 MTA2, it deacetylates p53 and modulates its effect on cell growth and apoptosis.

Protein: HDAC1 Gene: HDAC1 (Homo sapiens, chromosome 1, 32757708-32799224 [NCBI Reference Sequence: NC_(—)000001.10]; start site location: 32757771; strand: positive)

Gene Identification GeneID 3065 HGNC 4852 HPRD 03143 MIM 601241

Targeted Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 7075 CGCCTCCCGTCCCTACCGTCAGTCGGT 7 7141 CGGTCCGTCCGCCCTCCCGCCCGCGG 30 7207 CGCCAACTTGTGGTCCTACAGTCAACAAG 1740 7226 CGCAGACACGGGCCCGGAACTCGG 173 7258 CGCCCGGCCTAGGAGGGCAGGTTTCTC 1252

Target Shift Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 7075 CGCCTCCCGTCCCTACCGTCAGTCGGT 7 7076 GCCTCCCGTCCCTACCGTCA 8 7077 CCTCCCGTCCCTACCGTCAG 9 7078 CTCCCGTCCCTACCGTCAGT 10 7079 TCCCGTCCCTACCGTCAGTC 11 7080 CCCGTCCCTACCGTCAGTCG 12 7081 CCGTCCCTACCGTCAGTCGG 13 7082 CGTCCCTACCGTCAGTCGGT 14 7083 GTCCCTACCGTCAGTCGGTC 15 7084 TCCCTACCGTCAGTCGGTCC 16 7085 CCCTACCGTCAGTCGGTCCG 17 7086 CCTACCGTCAGTCGGTCCGT 18 7087 CTACCGTCAGTCGGTCCGTC 19 7088 TACCGTCAGTCGGTCCGTCC 20 7089 ACCGTCAGTCGGTCCGTCCG 21 7090 CCGTCAGTCGGTCCGTCCGC 22 7091 CGTCAGTCGGTCCGTCCGCC 23 7092 GTCAGTCGGTCCGTCCGCCC 24 7093 TCAGTCGGTCCGTCCGCCCT 25 7094 CAGTCGGTCCGTCCGCCCTC 26 7095 AGTCGGTCCGTCCGCCCTCC 27 7096 GTCGGTCCGTCCGCCCTCCC 28 7097 TCGGTCCGTCCGCCCTCCCG 29 7098 CGGTCCGTCCGCCCTCCCGC 30 7099 GGTCCGTCCGCCCTCCCGCC 31 7100 GTCCGTCCGCCCTCCCGCCC 32 7101 TCCGTCCGCCCTCCCGCCCG 33 7102 CCGTCCGCCCTCCCGCCCGC 34 7103 CGTCCGCCCTCCCGCCCGCG 35 7104 GTCCGCCCTCCCGCCCGCGG 36 7105 TCCGCCCTCCCGCCCGCGGC 37 7106 CCGCCCTCCCGCCCGCGGCT 38 7107 CGCCCTCCCGCCCGCGGCTC 39 7108 GCCCTCCCGCCCGCGGCTCC 40 7109 CCCTCCCGCCCGCGGCTCCG 41 7110 CCTCCCGCCCGCGGCTCCGC 42 7111 CTCCCGCCCGCGGCTCCGCT 43 7112 TCCCGCCCGCGGCTCCGCTC 44 7113 CCCGCCCGCGGCTCCGCTCA 45 7114 CCGCCCGCGGCTCCGCTCAG 46 7115 CGCCCGCGGCTCCGCTCAGC 47 7116 GCCCGCGGCTCCGCTCAGCG 48 7117 CCCGCGGCTCCGCTCAGCGT 49 7118 CCGCGGCTCCGCTCAGCGTC 50 7119 CGCGGCTCCGCTCAGCGTCC 51 7120 GCGGCTCCGCTCAGCGTCCG 52 7121 CGGCTCCGCTCAGCGTCCGA 53 7122 GGCTCCGCTCAGCGTCCGAC 54 7123 GCTCCGCTCAGCGTCCGACC 55 7124 CTCCGCTCAGCGTCCGACCC 56 7125 TCCGCTCAGCGTCCGACCCA 57 7126 CCGCTCAGCGTCCGACCCAG 58 7127 CGCTCAGCGTCCGACCCAGG 59 7128 GCTCAGCGTCCGACCCAGGG 60 7129 CTCAGCGTCCGACCCAGGGG 61 7130 TCAGCGTCCGACCCAGGGGG 62 7131 CAGCGTCCGACCCAGGGGGG 63 7132 AGCGTCCGACCCAGGGGGGA 64 7133 GCGTCCGACCCAGGGGGGAG 65 7134 CGTCCGACCCAGGGGGGAGG 66 7135 TCGCCTCCCGTCCCTACCGT 6 7136 CTCGCCTCCCGTCCCTACCG 5 7137 GCTCGCCTCCCGTCCCTACC 4 7138 TGCTCGCCTCCCGTCCCTAC 3 7139 TTGCTCGCCTCCCGTCCCTA 2 7140 CTTGCTCGCCTCCCGTCCCT 1 7141 CGGTCCGTCCGCCCTCCCGCCCGCGG 30 7142 GGTCCGTCCGCCCTCCCGCC 31 7143 GTCCGTCCGCCCTCCCGCCC 32 7144 TCCGTCCGCCCTCCCGCCCG 33 7145 CCGTCCGCCCTCCCGCCCGC 34 7146 CGTCCGCCCTCCCGCCCGCG 35 7147 GTCCGCCCTCCCGCCCGCGG 36 7148 TCCGCCCTCCCGCCCGCGGC 37 7149 CCGCCCTCCCGCCCGCGGCT 38 7150 CGCCCTCCCGCCCGCGGCTC 39 7151 GCCCTCCCGCCCGCGGCTCC 40 7152 CCCTCCCGCCCGCGGCTCCG 41 7153 CCTCCCGCCCGCGGCTCCGC 42 7154 CTCCCGCCCGCGGCTCCGCT 43 7155 TCCCGCCCGCGGCTCCGCTC 44 7156 CCCGCCCGCGGCTCCGCTCA 45 7157 CCGCCCGCGGCTCCGCTCAG 46 7158 CGCCCGCGGCTCCGCTCAGC 47 7159 GCCCGCGGCTCCGCTCAGCG 48 7160 CCCGCGGCTCCGCTCAGCGT 49 7161 CCGCGGCTCCGCTCAGCGTC 50 7162 CGCGGCTCCGCTCAGCGTCC 51 7163 GCGGCTCCGCTCAGCGTCCG 52 7164 CGGCTCCGCTCAGCGTCCGA 53 7165 GGCTCCGCTCAGCGTCCGAC 54 7166 GCTCCGCTCAGCGTCCGACC 55 7167 CTCCGCTCAGCGTCCGACCC 56 7168 TCCGCTCAGCGTCCGACCCA 57 7169 CCGCTCAGCGTCCGACCCAG 58 7170 CGCTCAGCGTCCGACCCAGG 59 7171 GCTCAGCGTCCGACCCAGGG 60 7172 CTCAGCGTCCGACCCAGGGG 61 7173 TCAGCGTCCGACCCAGGGGG 62 7174 CAGCGTCCGACCCAGGGGGG 63 7175 AGCGTCCGACCCAGGGGGGA 64 7176 GCGTCCGACCCAGGGGGGAG 65 7177 CGTCCGACCCAGGGGGGAGG 66 7178 TCGGTCCGTCCGCCCTCCCG 29 7179 GTCGGTCCGTCCGCCCTCCC 28 7180 AGTCGGTCCGTCCGCCCTCC 27 7181 CAGTCGGTCCGTCCGCCCTC 26 7182 TCAGTCGGTCCGTCCGCCCT 25 7183 GTCAGTCGGTCCGTCCGCCC 24 7184 CGTCAGTCGGTCCGTCCGCC 23 7185 CCGTCAGTCGGTCCGTCCGC 22 7186 ACCGTCAGTCGGTCCGTCCG 21 7187 TACCGTCAGTCGGTCCGTCC 20 7188 CTACCGTCAGTCGGTCCGTC 19 7189 CCTACCGTCAGTCGGTCCGT 18 7190 CCCTACCGTCAGTCGGTCCG 17 7191 TCCCTACCGTCAGTCGGTCC 16 7192 GTCCCTACCGTCAGTCGGTC 15 7193 CGTCCCTACCGTCAGTCGGT 14 7194 CCGTCCCTACCGTCAGTCGG 13 7195 CCCGTCCCTACCGTCAGTCG 12 7196 TCCCGTCCCTACCGTCAGTC 11 7197 CTCCCGTCCCTACCGTCAGT 10 7198 CCTCCCGTCCCTACCGTCAG 9 7199 GCCTCCCGTCCCTACCGTCA 8 7200 CGCCTCCCGTCCCTACCGTC 7 7201 TCGCCTCCCGTCCCTACCGT 6 7202 CTCGCCTCCCGTCCCTACCG 5 7203 GCTCGCCTCCCGTCCCTACC 4 7204 TGCTCGCCTCCCGTCCCTAC 3 7205 TTGCTCGCCTCCCGTCCCTA 2 7206 CTTGCTCGCCTCCCGTCCCT 1 7207 CGCCAACTTGTGGTCCTACAGTCAACAAG 1740 7208 CCGCCAACTTGTGGTCCTAC 1739 7209 GCCGCCAACTTGTGGTCCTA 1738 7210 AGCCGCCAACTTGTGGTCCT 1737 7211 TAGCCGCCAACTTGTGGTCC 1736 7212 TTAGCCGCCAACTTGTGGTC 1735 7213 GTTAGCCGCCAACTTGTGGT 1734 7214 AGTTAGCCGCCAACTTGTGG 1733 7215 AAGTTAGCCGCCAACTTGTG 1732 7216 TAAGTTAGCCGCCAACTTGT 1731 7217 CTAAGTTAGCCGCCAACTTG 1730 7218 TCTAAGTTAGCCGCCAACTT 1729 7219 CTCTAAGTTAGCCGCCAACT 1728 7220 GCTCTAAGTTAGCCGCCAAC 1727 7221 TGCTCTAAGTTAGCCGCCAA 1726 7222 TTGCTCTAAGTTAGCCGCCA 1725 7223 ATTGCTCTAAGTTAGCCGCC 1724 7224 CATTGCTCTAAGTTAGCCGC 1723 7225 ACATTGCTCTAAGTTAGCCG 1722 7226 CGCAGACACGGGCCCGGAACTCGG 173 7227 GCAGACACGGGCCCGGAACT 174 7228 CAGACACGGGCCCGGAACTC 175 7229 AGACACGGGCCCGGAACTCG 176 7230 GACACGGGCCCGGAACTCGG 177 7231 ACACGGGCCCGGAACTCGGC 178 7232 CACGGGCCCGGAACTCGGCA 179 7233 ACGGGCCCGGAACTCGGCAG 180 7234 CGGGCCCGGAACTCGGCAGG 181 7235 GGGCCCGGAACTCGGCAGGG 182 7236 GGCCCGGAACTCGGCAGGGG 183 7237 GCCCGGAACTCGGCAGGGGG 184 7238 CCCGGAACTCGGCAGGGGGC 185 7239 CCGGAACTCGGCAGGGGGCA 186 7240 GCGCAGACACGGGCCCGGAA 172 7241 TGCGCAGACACGGGCCCGGA 171 7242 TTGCGCAGACACGGGCCCGG 170 7243 CTTGCGCAGACACGGGCCCG 169 7244 GCTTGCGCAGACACGGGCCC 168 7245 AGCTTGCGCAGACACGGGCC 167 7246 CAGCTTGCGCAGACACGGGC 166 7247 TCAGCTTGCGCAGACACGGG 165 7248 ATCAGCTTGCGCAGACACGG 164 7249 AATCAGCTTGCGCAGACACG 163 7250 CAATCAGCTTGCGCAGACAC 162 7251 CCAATCAGCTTGCGCAGACA 161 7252 GCCAATCAGCTTGCGCAGAC 160 7253 AGCCAATCAGCTTGCGCAGA 159 7254 CAGCCAATCAGCTTGCGCAG 158 7255 CCAGCCAATCAGCTTGCGCA 157 7256 TCCAGCCAATCAGCTTGCGC 156 7257 CTCCAGCCAATCAGCTTGCG 155 7258 CGCCCGGCCTAGGAGGGCAGGTTTCTC 1252 7259 GCCCGGCCTAGGAGGGCAGG 1253 7260 CCCGGCCTAGGAGGGCAGGT 1254 7261 CCGGCCTAGGAGGGCAGGTT 1255 7262 CGGCCTAGGAGGGCAGGTTT 1256 7263 GCGCCCGGCCTAGGAGGGCA 1251 7264 AGCGCCCGGCCTAGGAGGGC 1250 7265 CAGCGCCCGGCCTAGGAGGG 1249 7266 ACAGCGCCCGGCCTAGGAGG 1248 7267 CACAGCGCCCGGCCTAGGAG 1247 7268 CCACAGCGCCCGGCCTAGGA 1246 7269 GCCACAGCGCCCGGCCTAGG 1245 7270 AGCCACAGCGCCCGGCCTAG 1244 7271 GAGCCACAGCGCCCGGCCTA 1243 7272 TGAGCCACAGCGCCCGGCCT 1242 7273 GTGAGCCACAGCGCCCGGCC 1241 7274 CGTGAGCCACAGCGCCCGGC 1240 7275 GCGTGAGCCACAGCGCCCGG 1239 7276 GGCGTGAGCCACAGCGCCCG 1238 7277 GGGCGTGAGCCACAGCGCCC 1237 7278 CGGGCGTGAGCCACAGCGCC 1236 7279 ACGGGCGTGAGCCACAGCGC 1235 7280 TACGGGCGTGAGCCACAGCG 1234 7281 TTACGGGCGTGAGCCACAGC 1233 7282 ATTACGGGCGTGAGCCACAG 1232 7283 GATTACGGGCGTGAGCCACA 1231 7284 AGATTACGGGCGTGAGCCAC 1230 7285 GAGATTACGGGCGTGAGCCA 1229 7286 TGAGATTACGGGCGTGAGCC 1228 7287 CTGAGATTACGGGCGTGAGC 1227 7288 GCTGAGATTACGGGCGTGAG 1226 7289 TGCTGAGATTACGGGCGTGA 1225 7290 ATGCTGAGATTACGGGCGTG 1224 7291 TATGCTGAGATTACGGGCGT 1223 7292 ATATGCTGAGATTACGGGCG 1222 7293 AATATGCTGAGATTACGGGC 1221 7294 CAATATGCTGAGATTACGGG 1220 7295 CCAATATGCTGAGATTACGG 1219 7296 CCCAATATGCTGAGATTACG 1218

Hot Zones (Relative upstream location to gene start site)  1-650  850-1300 1700-2050 2250-2550 2800-3700 4350-5000

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11976) CCAGGCTGATCTCAAACTCCTAAGCTCAAGTGATCCATGTTCCTCAGCCT CCCAAAGTGCTGGGATTATAGGCGTGAGCCATAGCGTCCAGCCCTGACTT ACATTTTAAAAGGATGGCTCTTGCTGCTGTCTTGAAAATAGACTGAGTTA GTCAGTTTATAAAACTGGGGAGATTTTGCATAAAACTCCAGATTTCTGCC TTCTCTTGAAAAATAGGGGCTAGGTGCGTTGGCTCACTCCTATAATCCCA GCATTTTGGAAGGCCAAGGTGGGCAGATTGCTTGAGCCCAGGAGTTTAAG ACCAGACTGGACAACATGGCAAAACCCTGTCTCTACCAAAAAAAAAAAAA AATTAGCAGGGTGTGGTGGTGCACACCTGCAGTCCCAGCTACTCAGGAGG CAAGCTTGTATTCCTAGCTACTTAGGAGGATAGTTTGAGCCCAGGAAGTC AAGGCTGCAGTGAGCATGATCCTGCCATTGCACTCCAGCCAGAGCAAAAA AGAGAGCGAAACCCCATCTCAAAAAAAAGGGAAGATTTAGCTATGTTGGA CTTACCTGTCCTCATGGAGCTGAATAATGGCCACCCCTCCAGGTAGGGCC TGAACTCTACCTTTGCCAGAGTCCCCTCCACTCCCTGTTGGTCTTAGACA ATGAAACTGAGTGTTAGTAGCTATTTACCACCAAGCTCATGCTTGTTGTT CTTATAATAAAGATAAATGGTTTAATAAATGGTATGATAAAGAAAATTAT ATTATGGTATTATACCATTTAATAAATGGTATAATAAAGAAAATGGTTTT TTGCACCCACATTTCCATTAAAAAGTGAGAAAATTAAAGATACCTGAGGA TGGCAGAGTGTTTGATGAAAGATAGGGAAATGTTGGCCAGGCACCGTGGC TCACACCTGTAATCCCAGCAGTTTAGGAGGCCGGGGCAGGCGGATCACAA GGTCAGGAGTTCAAGATCAGCCTGGCCAACATAGTGAAACCCCGTCTCTA CTAAAAATACAAAAAATTAGCCGGGAGTGGTGGCAGGTGCCTACAATCCC AGCTACTCGGAAGGCTGAATGGCGCGATCTCAGCTCATTGCAACCTCTGC CTCCCAGGTTCAAGCCATTCTTCTGCCTTAGCCTCCCTAGTAGCTGGGAT TACAGGCGCCTGCCACCATGCCTAGCTAATTTTTATATTTTTAGTAGACG TGGGGTTTCGCCATGTTGGCCAGGCTGGTCTCAAACTCCTGACCTTGGGT GATCTGCCCGCCTCGGTCTCCCAAACTGCTGGGATTACAGGAGTGAGCCA CAGTGCCCGGCCTCTAATTTTTATTTTTAATTTTTTTAATTTTTATTTTT TTAATTTTTATTTTATTTATTTTTTGTAATTTTTAAAATATACAAAAAAA GGGCCGGGTGTGGTGGCTCACGCCTGTAATCCCAGCACTTTTTGGGAGGC TGAGGTGGGTGGATCACGAGGTCAGGAGATCGAGACCATCCTGGCTAACA TGGTGAAACCCTGTCTCTACTAAAAATATAAAAAAATTAGCCGGGCCTGG TGGCAGGTGCCTGTAGTCCCAGCTACTCGGGAGGCTGAGACAGGAGAATG GCGTGAACTCGAGAGGTGGAGCTTGCAGTGAGCCAAGATCGCACCACTAC ACTCCAGCCTGGGCGACAGAGTGAGACTTCATCTCAAAAAAAAAAAAAAT TATATATATATATATACATATATATATGCAAACAAAGAGCATCTGAGTCA TAATAATGTAAATCTATCACCTGACTGACCTGCTGCCACACCTCATGATC TCATCTGATCCCCACACTCCTTCTCTTTGGGATACTGTGTACAGCCATAG CGTGGGTGAACTTTGTATTCCTATCCTCCCCATTTTTGTTATTTTATTTT ATTTCTTATTTATTTGAGACAGAGTCTCACTCTGTCATCCAGACTGTAAT GCAGTGGCCTGATCTCGGCTCACTGCAACCTCCACCTCCCGGTTTCAAGC GAATCTCCTGCCTCAGCCTCCTAAGTAGCTGGGACCTACAGGCACACACC ACCACGCCCAGCTAATTTTTGCATTTTTAATAGAGACGGGGTTTCACCGT GCTGGGCAGGCTGGTCTCGAACTCCTGACCTCAGGTGATTTGCCCACCTC AGCCTTCCAAATTGTTAGGATTACAGGCATGAGTCACTGTGCCCGGCCTC CTCCCCATTTTATAACAAGGGAAATGGAGGCCCAGAATGGTTAAGTAAAC CCACCCAGGGCTAGCTGAGAATTAGCAACAGAGAACTGGGAGTAGAATTT GTTCCCTGGCCCTTTGCTGTTTCTATTATAAGCCACCCAGTCTTAGATTT TCTGTTACCTTATAATTAATGACTCAAATGCAGTTTCTGAGTGAGAAACA CAAGTCCCAAACACTCTTTAAAGAGGCATAAAGATGTATCTTGTTGTTTT CTTTTGTTTGAGACAAGGCCTGGCTCTATTGCCCAGGCTGGAGTGTGGTG ACATGATCTTGGCTCACTGCAACATCTGTCTCCTGGGCTCAAGCCATCAT CCCACCTCAGCCTCCTGAGTAGCTGGAACTACAGGAGCGCGCCCCCACAC CTGGGTAATTTTTCTATTTTTTGTAGAGATGGGGTTTTGCCATGTTGCCC AGGCTGGTCTCGAACTCCTGAGCTCAAGTGATCCACCCATCTTGGCCTCC CAAAGTGCTGGGATTACAGGCGTGAGCCACTGTGCCCAGCCTCTTGTTGA CTGTAGGACCACAAGTTGGCGGCTAACTTAGAGCAATGTTTGGCACACAG GAAGCACTCATTAAATATTGACATTATTGTAGTTATTTTAATAGCCCAGC ATTGCACTTTTAGGTCTTTCAGCTTTCAGTGATGATCAGTTGATAATTGA TGATCTGGTGGAGTGGTTCTTAATGGTAGAGTTGGGGGCAATTTTACACT CCCTTACACCCCACTAATCTTCCCCCCAACCCAATGGTAAAGCTATTGCA CAGTACTTGGCAATGTCTAGAGACAATTTTGGTTGTCACAGCCTGGGGGG AAGGTGCTACTGGCATCTAGTGGGTAGAGGCTAAGGATGCTGCTTAATTT TTTTTTTTTTTGAGACAAGAGTTTCACTCTAGTTGCCCAGGCACAGAACA GCTTCACAGAAGCTGTTAATGCACAGAATAGCTTCCTACAAAAAAGCATT ACCTGGCCCAAAATGTCATTAGCTACCAGGCTGAGAAACCTGCCCTCCTA GGCCGGGCGCTGTGGCTCACGCCCGTAATCTCAGCATATTGGGAGGCCGA GGTGGGCGGATCCTGAGGTCAGGAGTTCGAGACCACCTGGACCAACACGG AGAAACCCAGTCTCTACCAAAAATACAAAATTAGCCGGGCATGGTGGCAC ATGCTTGTAATCCCAGCTACTCGGGAGGCTGAGGCAGGAGAACCGCTTGA ACACAGAGGCAGAGATTGTGGTGAGCCGAGATCACACCATTGCACTCCAG CCTGGGCAACTAGAGCGAAACTCTTGTCTCAAAAAAAAAAAAAAAAAAAA AAACAGGGAAAGAAAAGAAAGGAAACCTGCCCTCCTATCATAGGATAATC CCATTTCCTCCTGTCTAAAGAGACGCCTACTTAGTCATCCTGGGTGACTG CATCAGGGAGGTAGATTTTGGAGTCTGAAAGGCTGGGTTCTGTCACTTTG TTACAGTGCCTCTGGTGCAAAGAAAGCATTTTAAAAACCCTGTACAATTA AAAAATTGAATTTAATTACTTTGTGTAACTTTGAATAATTCACAGAAGTC TGAACTTCTTTATCCTGTCCTGTAAAATGGAGGTAAAAAGCCCTTGGCCG AGAGCTGTTTTGAGGAAAAACTGAAATAACATTGGTAAAGTGTCGCACAG TACTTGGCACACAGCAGCCCCTCGACAAACATTAGCTTTCTTTCCCTTTC TTGTCGGTTTCTTCCTCTCCAAACCCGCGTGTTGCTTTTCTTTTTAATTA TTTTTCTGTAGCCCTCCTTTGCGGCCACAAACTCGCTTTCTAACCCAGGT TCAGCCCTTTTATTGGCTGAGTGACCTTGTGCAAGTCACTTTTCCCCTGT AGGCCTCGGTTTATTCTCCGTAAAATCAGAAAGTTGGCCTCCGATCTCCA AGCACGCTTTTCACGACGAAGTGGGACTGTTAAGTTTACAGAGCTGCTTT CCTCCCCCGGGACTGATGGTACGGTCCCCGGGCGGCTCCCCACCCATCTG TCGCAGACCTTGGTACAGGCCCAGGGGGCCCTCGGCGGCCTCTCCGGGCT GCCCTTGCCCCCTGCCGAGTTCCGGGCCCGTGTCTGCGCAAGCTGATTGG CTGGAGCGGTGCCCGGGCTGCGCGGCTATAGGTGAGCCCAGGAGGGGACG GGCGGGGCGGGCCGGAGGCCCGCCCCCTCCCCCCTGGGTCGGACGCTGAG CGGAGCCGCGGGCGGGAGGGCGGACGGACCGACTGACGGTAGGGACGGGA GGCGAGCAAG ATG

28. PD-1. Programmed cell death protein 1 (PD-1) is also known as CD279 (cluster of differentiation 279). This gene encodes a cell surface membrane protein of the immunoglobulin superfamily. This protein is expressed in pro-B cells and is thought to play a role in their differentiation. PD-1 has two ligands, PD-L1 and PD-L2. PD-L1 protein is upregulated on macrophages and dendritic cells (DC) in response to LPS and GM-CSF treatment, and on T cells and B cells upon TCR and B cell receptor signaling.

Monoclonal antibodies blocking PD-1 may overcome immune resistance and boost the immune system are being developed for the treatment of cancer (Weber 2010, Semin. Oncol. 37 (5): 430-9). Nivolumab, a representative antibody, produced complete or partial responses in non-small-cell lung cancer, melanoma, and renal-cell cancer, in a clinical trial with a total of 296 patients; colon and pancreatic cancer did not have a response (Topalian et al., 2012: N Engl J Med 2012; 366:2443-2454). In HIV, drugs targeting PD-1 may augment immune responses and/or facilitate HIV eradication.

Protein: PD-1 Gene: PDCD1 (Homo sapiens, chromosome 2, 242792033-242801058 [NCBI Reference Sequence: NC_(—)000002.11]; start site location: 242800990; strand: negative)

Gene Identification GeneID 5133 HGNC 8760 HPRD 02590 MIM 600244

Targeted Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 7297 TGCCGCCTTCTCCACTGCTCAGGCG 23 7316 ACCGCCTGACAGCTGGCGCGGCTGCCTGGC 1061 7379 CTGCGAGGCGCGGCCACGGCG 1171 7396 CGAGGAGGAAAGGCAGGCGGAGTCCG 3395 7397 CAGCGAAGCTGCAGAACGTCCCCATCACCACG 4268 7439 CGACAGCCGTGGGAAGGTGCAGTACG 4388 7440 CGGGATTCCCTGGAGATGCCTCCAGCGCG 4422 7466 AGGCGGTCCCAGGGCTCAGGTGTGGG 2229 7498 GCGTGCACCCCGTGGCCAGCTC 3813 7526 CAACGTACACGCAATCCACAAC 2832

Target Shift Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 7297 TGCCGCCTTCTCCACTGCTCAGGCG 23 7298 GCCGCCTTCTCCACTGCTCA 24 7299 CCGCCTTCTCCACTGCTCAG 25 7300 CGCCTTCTCCACTGCTCAGG 26 7301 GTGCCGCCTTCTCCACTGCT 22 7302 AGTGCCGCCTTCTCCACTGC 21 7303 GAGTGCCGCCTTCTCCACTG 20 7304 AGAGTGCCGCCTTCTCCACT 19 7305 CAGAGTGCCGCCTTCTCCAC 18 7306 CCAGAGTGCCGCCTTCTCCA 17 7307 ACCAGAGTGCCGCCTTCTCC 16 7308 CACCAGAGTGCCGCCTTCTC 15 7309 CCACCAGAGTGCCGCCTTCT 14 7310 CCCACCAGAGTGCCGCCTTC 13 7311 CCCCACCAGAGTGCCGCCTT 12 7312 GCCCCACCAGAGTGCCGCCT 11 7313 AGCCCCACCAGAGTGCCGCC 10 7314 CAGCCCCACCAGAGTGCCGC 9 7315 GCAGCCCCACCAGAGTGCCG 8 7316 ACCGCCTGACAGCTGGCGCGGCTGCCTGGC 1061 7317 CCGCCTGACAGCTGGCGCGG 1062 7318 CGCCTGACAGCTGGCGCGGC 1063 7319 GCCTGACAGCTGGCGCGGCT 1064 7320 CCTGACAGCTGGCGCGGCTG 1065 7321 CTGACAGCTGGCGCGGCTGC 1066 7322 TGACAGCTGGCGCGGCTGCC 1067 7323 GACAGCTGGCGCGGCTGCCT 1068 7324 ACAGCTGGCGCGGCTGCCTG 1069 7325 CAGCTGGCGCGGCTGCCTGG 1070 7326 AGCTGGCGCGGCTGCCTGGC 1071 7327 GCTGGCGCGGCTGCCTGGCT 1072 7328 CTGGCGCGGCTGCCTGGCTC 1073 7329 TGGCGCGGCTGCCTGGCTCC 1074 7330 GGCGCGGCTGCCTGGCTCCG 1075 7331 GCGCGGCTGCCTGGCTCCGA 1076 7332 CGCGGCTGCCTGGCTCCGAG 1077 7333 GCGGCTGCCTGGCTCCGAGA 1078 7334 CGGCTGCCTGGCTCCGAGAG 1079 7335 GGCTGCCTGGCTCCGAGAGA 1080 7336 GCTGCCTGGCTCCGAGAGAC 1081 7337 CTGCCTGGCTCCGAGAGACA 1082 7338 TGCCTGGCTCCGAGAGACAC 1083 7339 GCCTGGCTCCGAGAGACACT 1084 7340 CCTGGCTCCGAGAGACACTC 1085 7341 CTGGCTCCGAGAGACACTCG 1086 7342 TGGCTCCGAGAGACACTCGG 1087 7343 GGCTCCGAGAGACACTCGGC 1088 7344 GCTCCGAGAGACACTCGGCC 1089 7345 CTCCGAGAGACACTCGGCCC 1090 7346 TCCGAGAGACACTCGGCCCG 1091 7347 CCGAGAGACACTCGGCCCGG 1092 7348 CGAGAGACACTCGGCCCGGC 1093 7349 GAGAGACACTCGGCCCGGCT 1094 7350 AGAGACACTCGGCCCGGCTC 1095 7351 GAGACACTCGGCCCGGCTCT 1096 7352 AGACACTCGGCCCGGCTCTG 1097 7353 GACACTCGGCCCGGCTCTGA 1098 7354 ACACTCGGCCCGGCTCTGAA 1099 7355 CACTCGGCCCGGCTCTGAAG 1100 7356 ACTCGGCCCGGCTCTGAAGG 1101 7357 CTCGGCCCGGCTCTGAAGGG 1102 7358 TCGGCCCGGCTCTGAAGGGA 1103 7359 CGGCCCGGCTCTGAAGGGAA 1104 7360 GGCCCGGCTCTGAAGGGAAA 1105 7361 GCCCGGCTCTGAAGGGAAAA 1106 7362 CCCGGCTCTGAAGGGAAAAC 1107 7363 CCGGCTCTGAAGGGAAAACA 1108 7364 CGGCTCTGAAGGGAAAACAT 1109 7365 AACCGCCTGACAGCTGGCGC 1060 7366 AAACCGCCTGACAGCTGGCG 1059 7367 GAAACCGCCTGACAGCTGGC 1058 7368 AGAAACCGCCTGACAGCTGG 1057 7369 TAGAAACCGCCTGACAGCTG 1056 7370 CTAGAAACCGCCTGACAGCT 1055 7371 GCTAGAAACCGCCTGACAGC 1054 7372 GGCTAGAAACCGCCTGACAG 1053 7373 AGGCTAGAAACCGCCTGACA 1052 7374 GAGGCTAGAAACCGCCTGAC 1051 7375 CGAGGCTAGAAACCGCCTGA 1050 7376 GCGAGGCTAGAAACCGCCTG 1049 7377 AGCGAGGCTAGAAACCGCCT 1048 7378 AAGCGAGGCTAGAAACCGCC 1047 7379 CTGCGAGGCGCGGCCACGGCG 1171 7380 TGCGAGGCGCGGCCACGGCG 1172 7381 GCGAGGCGCGGCCACGGCGA 1173 7382 CGAGGCGCGGCCACGGCGAG 1174 7383 TCTGCGAGGCGCGGCCACGG 1170 7384 GTCTGCGAGGCGCGGCCACG 1169 7385 TGTCTGCGAGGCGCGGCCAC 1168 7386 ATGTCTGCGAGGCGCGGCCA 1167 7387 GATGTCTGCGAGGCGCGGCC 1166 7388 TGATGTCTGCGAGGCGCGGC 1165 7389 ATGATGTCTGCGAGGCGCGG 1164 7390 GATGATGTCTGCGAGGCGCG 1163 7391 AGATGATGTCTGCGAGGCGC 1162 7392 AAGATGATGTCTGCGAGGCG 1161 7393 AAAGATGATGTCTGCGAGGC 1160 7394 CAAAGATGATGTCTGCGAGG 1159 7395 TCAAAGATGATGTCTGCGAG 1158 7396 CGAGGAGGAAAGGCAGGCGGAGTCCG 3395 7397 CAGCGAAGCTGCAGAACGTCCCCATCACCACG 4268 7398 AGCGAAGCTGCAGAACGTCC 4269 7399 GCGAAGCTGCAGAACGTCCC 4270 7400 CGAAGCTGCAGAACGTCCCC 4271 7401 GAAGCTGCAGAACGTCCCCA 4272 7402 AAGCTGCAGAACGTCCCCAT 4273 7403 AGCTGCAGAACGTCCCCATC 4274 7404 GCTGCAGAACGTCCCCATCA 4275 7405 CTGCAGAACGTCCCCATCAC 4276 7406 TGCAGAACGTCCCCATCACC 4277 7407 GCAGAACGTCCCCATCACCA 4278 7408 CAGAACGTCCCCATCACCAC 4279 7409 AGAACGTCCCCATCACCACG 4280 7410 GAACGTCCCCATCACCACGG 4281 7411 AACGTCCCCATCACCACGGG 4282 7412 ACGTCCCCATCACCACGGGG 4283 7413 CGTCCCCATCACCACGGGGT 4284 7414 GTCCCCATCACCACGGGGTC 4285 7415 TCCCCATCACCACGGGGTCC 4286 7416 CCCCATCACCACGGGGTCCT 4287 7417 CCCATCACCACGGGGTCCTC 4288 7418 CCATCACCACGGGGTCCTCC 4289 7419 CATCACCACGGGGTCCTCCG 4290 7420 ATCACCACGGGGTCCTCCGG 4291 7421 TCACCACGGGGTCCTCCGGG 4292 7422 CACCACGGGGTCCTCCGGGT 4293 7423 ACCACGGGGTCCTCCGGGTG 4294 7424 CCACGGGGTCCTCCGGGTGC 4295 7425 CACGGGGTCCTCCGGGTGCC 4296 7426 ACGGGGTCCTCCGGGTGCCC 4297 7427 CGGGGTCCTCCGGGTGCCCT 4298 7428 GGGGTCCTCCGGGTGCCCTT 4299 7429 GGGTCCTCCGGGTGCCCTTG 4300 7430 GGTCCTCCGGGTGCCCTTGG 4301 7431 GTCCTCCGGGTGCCCTTGGC 4302 7432 TCCTCCGGGTGCCCTTGGCA 4303 7433 CCTCCGGGTGCCCTTGGCAA 4304 7434 CTCCGGGTGCCCTTGGCAAT 4305 7435 TCCGGGTGCCCTTGGCAATA 4306 7436 CCGGGTGCCCTTGGCAATAC 4307 7437 CGGGTGCCCTTGGCAATACA 4308 7438 ACAGCGAAGCTGCAGAACGT 4267 7439 CGACAGCCGTGGGAAGGTGCAGTACG 4388 7440 CGGGATTCCCTGGAGATGCCTCCAGCGCG 4422 7441 CCGGGATTCCCTGGAGATGC 4421 7442 TCCGGGATTCCCTGGAGATG 4420 7443 TTCCGGGATTCCCTGGAGAT 4419 7444 CTTCCGGGATTCCCTGGAGA 4418 7445 CCTTCCGGGATTCCCTGGAG 4417 7446 TCCTTCCGGGATTCCCTGGA 4416 7447 ATCCTTCCGGGATTCCCTGG 4415 7448 CATCCTTCCGGGATTCCCTG 4414 7449 GCATCCTTCCGGGATTCCCT 4413 7450 CGCATCCTTCCGGGATTCCC 4412 7451 ACGCATCCTTCCGGGATTCC 4411 7452 TACGCATCCTTCCGGGATTC 4410 7453 GTACGCATCCTTCCGGGATT 4409 7454 AGTACGCATCCTTCCGGGAT 4408 7455 CAGTACGCATCCTTCCGGGA 4407 7456 GCAGTACGCATCCTTCCGGG 4406 7457 TGCAGTACGCATCCTTCCGG 4405 7458 GTGCAGTACGCATCCTTCCG 4404 7459 GGTGCAGTACGCATCCTTCC 4403 7460 AGGTGCAGTACGCATCCTTC 4402 7461 AAGGTGCAGTACGCATCCTT 4401 7462 GAAGGTGCAGTACGCATCCT 4400 7463 GGAAGGTGCAGTACGCATCC 4399 7464 GGGAAGGTGCAGTACGCATC 4398 7465 TGGGAAGGTGCAGTACGCAT 4397 7466 AGGCGGTCCCAGGGCTCAGGTGTGGG 2229 7467 GGCGGTCCCAGGGCTCAGGT 2230 7468 GCGGTCCCAGGGCTCAGGTG 2231 7469 CGGTCCCAGGGCTCAGGTGT 2232 7470 TAGGCGGTCCCAGGGCTCAG 2228 7471 ATAGGCGGTCCCAGGGCTCA 2227 7472 GATAGGCGGTCCCAGGGCTC 2226 7473 AGATAGGCGGTCCCAGGGCT 2225 7474 CAGATAGGCGGTCCCAGGGC 2224 7475 GCAGATAGGCGGTCCCAGGG 2223 7476 AGCAGATAGGCGGTCCCAGG 2222 7477 AAGCAGATAGGCGGTCCCAG 2221 7478 GAAGCAGATAGGCGGTCCCA 2220 7479 CGAAGCAGATAGGCGGTCCC 2219 7480 CCGAAGCAGATAGGCGGTCC 2218 7481 CCCGAAGCAGATAGGCGGTC 2217 7482 CCCCGAAGCAGATAGGCGGT 2216 7483 ACCCCGAAGCAGATAGGCGG 2215 7484 CACCCCGAAGCAGATAGGCG 2214 7485 CCACCCCGAAGCAGATAGGC 2213 7486 CCCACCCCGAAGCAGATAGG 2212 7487 CCCCACCCCGAAGCAGATAG 2211 7488 ACCCCACCCCGAAGCAGATA 2210 7489 GACCCCACCCCGAAGCAGAT 2209 7490 GGACCCCACCCCGAAGCAGA 2208 7491 GGGACCCCACCCCGAAGCAG 2207 7492 TGGGACCCCACCCCGAAGCA 2206 7493 CTGGGACCCCACCCCGAAGC 2205 7494 CCTGGGACCCCACCCCGAAG 2204 7495 TCCTGGGACCCCACCCCGAA 2203 7496 GTCCTGGGACCCCACCCCGA 2202 7497 GGTCCTGGGACCCCACCCCG 2201 7498 GCGTGCACCCCGTGGCCAGCTC 3813 7499 CGTGCACCCCGTGGCCAGCT 3814 7500 GTGCACCCCGTGGCCAGCTC 3815 7501 TGCACCCCGTGGCCAGCTCA 3816 7502 GCACCCCGTGGCCAGCTCAT 3817 7503 CACCCCGTGGCCAGCTCATA 3818 7504 ACCCCGTGGCCAGCTCATAT 3819 7505 CCCCGTGGCCAGCTCATATC 3820 7506 CCCGTGGCCAGCTCATATCT 3821 7507 CCGTGGCCAGCTCATATCTA 3822 7508 CGTGGCCAGCTCATATCTAA 3823 7509 GGCGTGCACCCCGTGGCCAG 3812 7510 AGGCGTGCACCCCGTGGCCA 3811 7511 CAGGCGTGCACCCCGTGGCC 3810 7512 ACAGGCGTGCACCCCGTGGC 3809 7513 CACAGGCGTGCACCCCGTGG 3808 7514 CCACAGGCGTGCACCCCGTG 3807 7515 ACCACAGGCGTGCACCCCGT 3806 7516 GACCACAGGCGTGCACCCCG 3805 7517 GGACCACAGGCGTGCACCCC 3804 7518 GGGACCACAGGCGTGCACCC 3803 7519 TGGGACCACAGGCGTGCACC 3802 7520 CTGGGACCACAGGCGTGCAC 3801 7521 GCTGGGACCACAGGCGTGCA 3800 7522 AGCTGGGACCACAGGCGTGC 3799 7523 TAGCTGGGACCACAGGCGTG 3798 7524 GTAGCTGGGACCACAGGCGT 3797 7525 AGTAGCTGGGACCACAGGCG 3796 7526 CAACGTACACGCAATCCACAAC 2832 7527 AACGTACACGCAATCCACAA 2833 7528 ACGTACACGCAATCCACAAC 2834 7529 CGTACACGCAATCCACAACA 2835 7530 GTACACGCAATCCACAACAC 2836 7531 TACACGCAATCCACAACACA 2837 7532 ACACGCAATCCACAACACAT 2838 7533 CACGCAATCCACAACACATA 2839 7534 ACGCAATCCACAACACATAC 2840 7535 CGCAATCCACAACACATACA 2841 7536 CCAACGTACACGCAATCCAC 2831 7537 CCCAACGTACACGCAATCCA 2830 7538 CCCCAACGTACACGCAATCC 2829 7539 TCCCCAACGTACACGCAATC 2828 7540 ATCCCCAACGTACACGCAAT 2827 7541 AATCCCCAACGTACACGCAA 2826 7542 CAATCCCCAACGTACACGCA 2825 7543 ACAATCCCCAACGTACACGC 2824 7544 CACAATCCCCAACGTACACG 2823 7545 GCACAATCCCCAACGTACAC 2822 7546 TGCACAATCCCCAACGTACA 2821 7547 ATGCACAATCCCCAACGTAC 2820 7548 CATGCACAATCCCCAACGTA 2819 7549 ACATGCACAATCCCCAACGT 2818 7550 AACATGCACAATCCCCAACG 2817

Hot Zones (Relative upstream location to gene start site)   1-1450 1850-2350 2750-3000 3100-3600 3650-4050 4100-5000

Examples

In FIG. 48, In MCF7 (human mammary breast cell line), PD1 (293) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The PD1 sequence PD1 (293) fits the independent and dependent DNAi motif claims.

The secondary structure for PD1 (293) is shown in FIG. 49.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11977) ACCACAGCCTGGATGGCTCGGCACAGAGAGGAACGGGCTGCTGAAACACA CGCGCTGGAGGCATCTCCAGGGAATCCCGGAAGGATGCGTACTGCACCTT CCCACGGCTGTCGCTGGAATGAAACAGTGATGGGGGTGGAGGGCAGGCTC GTGGCCACCAGCAGGGAGGTGGGTGTATTGCCAAGGGCACCCGGAGGACC CCGTGGTGATGGGGACGTTCTGCAGCTTCGCTGTCAGGGGATGCAGGAGC CTTCGCTTGTGCCCACATTGTGCAGCCTTTGATAGGCACACATTAGCCAG AAACGGGGACTCAGGATGGGATCGAGGTGTCACATCAAAGTCATTAACCT GGTTGTGACGTTGTCCTGTGGTTTTCCAAACTGTTATCATTCAGAGAGAC TGAGCAGAGTGTATGAGGAAACATCTGTGTAATTTCTTACAACTGCAAGT AAATCTACAATTATCTCAATTGTAAATGATACAATACTCAACCAAAACAT ACAACCATCAGCCAGGTGTGGTGGCTCACGCCTGTAATCCCAACTCTTTG GGAGTCCAAGGTGGGAGAATTGCTGGAGGCCCGGAGTTTGAGACCAGCCT GGGTAACATAAAGAGACCTCCTCTGCCCCCACCCCAAATTCTACAAAAAA AAAAAATGTTAGATATGAGCTGGCCACGGGGTGCACGCCTGTGGTCCCAG CTACTCAGGAGGCTGAGGTCGGAGGATCGCTTGAGCCCAGGAGGTCGAGG CTGCAGTGAGCCAAGATCACACCACTGCACTCCAGCCTGGGTGCAGAGCA AGACCCTGTCTCTAAAAGAAAATAAACAGACAAAAACCACATACAACTTT GCTTGTTGTAAATTATCTTTTAACTGAATGCCCTGGATTGAATCTGGCTG CTGCCATCCCAGGGCCAGTGATTTGGATGGGGTATGACCCTCTGTGAGGA AGGAGCAGGCGGTGGGGGAAGGGCCTGGGTGTCCAGGTTCCCTGGGAAGG AAGGCTGAGAAAAGGAGATGGGGGAGGGGTGCGCAGGGCCGGCCAGCCAA GGGCCCCTTAGCCCCATCTACCCTGCTCCCCGGACTCCGCCTGCCTTTCC TCCTCGTGACAGAAGACAGTGGAAGCCTACTGGGTGGAAGGCACGGGCTT AGGATGTGTGTGGGAGGAAAGTGTGTGTGCTGGGGAGCATGTATGTTTGG GAGTTGTGTGTGTTGGAAATCGTGTGTTGGGGATTGTGTGTATATTGCAG ATTTTGTATGTGTGTTGGGGATTGTGGTGTGTGGGTGTTGTAGATTGCGT GTTGGGGATTGTGTTGGGGATTGTGTATGTGTTGGGGGTTGTGTGTGTGT TGGGGATTGTGTGTGGGGGAGATTGTGTGTGTGTGCTGGGGATTGGGTGT GTTGGGGATTGTGTGTGTGTTGAGGATTGTGTGTGGGGGAGATTCTGTGT GTGTGCTGGGGATTGGGTGTGTTGGGGATTGTGTGTGTATTGGGAATTGT GTGTGTGTTGAGGATTGTGTGTGTTGGGGATTTGTGTGTGTGTTGGGGAT TCTGTGCATGTTGGGAGTTGTGTGTGAGTTGGGGACAATGTGTACAGAGG ATTGTGTGTTGGAAATTTTGTGTGTGCGTTGGGAATTTTGTGTATGTGTT GTGGATTGCGTGTACGTTGGGGATTGTGCATGTTGGGAATTTTGTGTGTG TGTTGAGAATTGTGTGTGAGGGAATTGTGTGTGTTTGAGATTGTGTGTGT ATTGGGAATTGTGTGTGTGTTGAGGATTGTGTGTGTTCTGAGGATTGTAT GTGTTGGGAATTTTGTGTGTGTGTTGAGGATTGTGTGTGTTGGGGATTCT ACGTATGTTGAAAGTTGTGTGTGTGTTGGGATTGTGTGTGTGTTGTGGAT TGTGTGTGTTGGGAATTGTGTGTGTGTGTTGAGGATTGTGTGCAGGGGGA TTGTGTGTGTTGGAGATTGTATGTGTTGGGAATTTTGTGTGTGTGTTGGG GACTGTGTATGTTTTGGGGATTGTGTGTGTTGGGAATTTTGTGTGTGTGT TGAGGATTGTGTGTGGGGGGATTGTGTGTGTTGGAGATTGTGTGTGTGTT GGGGACTGTGTGTGTGTTGGGGACTGTGTGTGTTGGGGTGTGGTGTGTTG GAAATCGTGTGTTGGGGACACCGTATGTGTTTGGGGGAGGGTGTCAATAA GTGGTCTGGAGTGTGATATTGGGGTGCAGGCTCCATGAGTCCCCACCCCA CACCTGAGCCCTGGGACCGCCTATCTGCTTCGGGGTGGGGTCCCAGGACC CTGTAGGTTCAGCCTACTAGTCCAGGCCCAATGCCCAATGCCTGCATCCC TGCAGGCCCTGTGCTCTCCAGGCTCAGACCCCTCGCAGCCCTGCAGACCC TCCCTGGGTCCATGTGTCTCTTTGCAGGTGCTCCAGCGAGTAGCAATGTG GAGAGACCATCAGGCAGCCCTGGCCTCAGTGGCCGCAGTCCCCTGGCTCC ACGCTGGGCCCACCCCACCAGGTCTCCTCTCCCATGGCCCAGGGGCCTTC AGTGGGACTGAGAGGAGGAGGGAAGGAGAGTGGGTGACAGGGAAGAACTG CAGGGAGAGAGGAGAGGGGTGGGAGAAGGAGAAGGAAGGAAGGGGTAGGA TGGAAGCTGGGTTTCTCCCTGTGCCCGCCCCCTACTCCAGGACATGTGTC CAAGCCCTGGCAGGTGGAATTTTGGGGGCAGGGCCTTGGTGGTGAGGAGA CCTTCCAGGGGTCTGATAGCATCTCCCATCTCAGAGCCCACCTCCTGGGC CCAGCCTCCCCTCCAGCCCACACAGTGGCATTCCCAGTCCTCAGAGGACA GCTTCGTCCCACAAAGCTCAGAGCCTTGAGGAAGGCCCACTGCTGCCCTG GAACAGAGACAGCATTCAACAGAGGTTGGAACAAGGCTCTACAGGGCTGG GGGCAGAGGGAGGTTCTGTCCAGAATCTGCCTTCAGGACAAGTACAGCCA GCAGGGGCAGCTTAGCCACTTATCCACTGCCTGGGCGAGGCACAGGGCTA TGGAGGCACCTACCAACCAACAGTTCTCCAGCCCCAGAGCCCCAGCCCCT GAGGCACAAGGGTGGGTGTGCCAGGAGACAGTTGCTGCGGGCCACCTTAG CTGTCTGGCAGCACAGTGGGTGCTGCCAGGCTCCCTGGGGGCCCCCCGCC AAGCCCACCTGGCCAGCTGGGCCCCCCCCACCTCCCCACCAAGCCACCCA CACAGCCTCACATCTCTGAGACCCGGGAGTGGCCCTTTGTTCATAAACGA GAGCTTCCTCGCCGTGGCCGCGCCTCGCAGACATCATCTTTGATGCTCTT TTTCCACTGTTTCGGTGCTTTAATGTTTTCCCTTCAGAGCCGGGCCGAGT GTCTCTCGGAGCCAGGCAGCCGCGCCAGCTGTCAGGCGGTTTCTAGCCTC GCTTCGGTTATTTTAAGCTGATGAGCCTGACGCATCTCATCACTAATATC AGCAGTTTCATTTCTCCTGTTTTCCATTCGCTGTAATAAAATGCTCAGCA CAGAATACAAGGAGATAAGCAAGCCATTTCACAAACGCCGGGCCGCCAGC CAGGCCCAGGCACTGGACCCCCTGAACCACCCCACCCTGGCACGAGTGGG CTGGAGGGCAGGGCCCCGGGGAAGAAGGTCAAGGCTGGAAGGGGAGGTCA GCCTCACAGCCAGCCCCTGCCACCGCCCCAGCCCCCCCGTCAGGCTGTTG CAGGCATCACACGGTGGAAAGATCTGGAACTGTGGCCATGGTGTGAGGCC ATCCACAAGGTGGAAGCTTTGAGGGGGAGCCGATTAGCCATGGACAGTTG TCATTCAGTAGGGTCACCTGTGCCCCAGCGAAGGGGGATGGGCCGGGAAG GCAGAGGCCAGGCACCTGCCCCCAGCAGGGGCAGAGGCTGTGGGCAGCCG GGAGGCTCCCAGAGGCTCCGACAGAATGGGAGTGGGGTTGAGCCCACCCC TCACTGCAGCCCAGGAACCTGAGCCCAGAGGGGGCCACCCACCTTCCCCA GGCAGGGAGGCCCGGCCCCCAGGGAGATGGGGGGGATGGGGGAGGAGAAG GGCCTGCCCCCACCCGGCAGCCTCAGGAGGGGCAGCTCGGGCGGGATATG GAAAGAGGCCACAGCAGTGAGCAGAGACACAGAGGAGGAAGGGGCCCTGA GCTGGGGAGACCCCCACGGGGTAGGGCGTGGGGGCCACGGGCCCACCTCC TCCCCATCTCCTCTGTCTCCCTGTCTCTGTCTCTCTCTCCCTCCCCCACC CTCTCCCCAGTCCTACCCCCTCCTCACCCCTCCTCCCCCAGCACTGCCTC TGTCACTCTCGCCCACGTGGATGTGGAGGAAGAGGGGGCGGGAGCAAGGG GCGGGCACCCTCCCTTCAACCTGACCTGGGACAGTTTCCCTTCCGCTCAC CTCCGCCTGAGCAGTGGAGAAGGCGGCACTCTGGTGGGGCTGCTCCAGGC ATG

29. BCL2. Bcl-2 (B-cell lymphoma 2) is the founding member of the Bcl-2 family of apoptosis regulator proteins encoded by the BCL2 gene that was first described in chromosomal translocations involving chromosomes 14 and 18 in follicular lymphomas (Tsujimoto et al. Science 226 (4678): 1097-99). The dysregulation of cell death is a defining characteristic of malignant cells and BCL-2 protein plays a key and central role. BCL-2 confers an anti-apoptotic phenotype that contributes to the genesis of hematopoietic and lymphatic cancers. In many cases of diffuse large B-cell (DLBCL) and follicular lymphomas (FL), BCL2 overexpression is driven by the t(14,18) chromosomal rearrangement of the BCL2 oncogene. In chronic lymphocytic leukemia, impaired degradation of BCL2 mRNA causes continuous production of BCL2. The Bcl-2 gene has been implicated in a number of cancers, including melanoma, breast, prostate, chronic lymphocytic leukemia, skin, sarcoma, and lung carcinomas, as well as schizophrenia and autoimmunity. It is also thought to be involved in resistance to conventional cancer treatment and evidence also suggests that decreased apoptosis may play a role in the development of cancer.

Protein: BCL2 Gene: BCL2 (Homo sapiens, chromosome 18, 63123346-63319778 [NCBI Reference Sequence: NC_(—)000018.10]; start site location: 63318666; strand: negative)

Gene Identification GeneID 596 HGNC 990 MIM 151430

Targeted Sequences Relative upstream location to Sequence Design gene start ID ID Sequence (5′-3′) site 13682 TGTCCACCTGAACACCTAGTCC 2388

In FIG. 50, In MDA-MB-231 (human breast cell line), BL2 at 10 μM showed increased inhibition compared to BL3 and BL4 (10 μM). The BL2 (structure shown below) fits the independent and dependent DNAi motif claims. Both BL3 and BL4 contained a single mismatched base meaning neither sequence had 100% homology to its complementary strand. This demonstrates that many times even a single mismatch to the complementary strand decreases the inhibitory effects of a DNAi oligonucleotide. The mismatches for BL3 and BL4 are noted below with the mismatched letter highlighted and bolded. It should also be noted that a 20-mer version of BL2 demonstrated similar significant inhibition (data not shown) as the 24-mer version of BL2 shown in FIGS. 50, 51, and 52.

In FIG. 51, M14 (human melanoma cell line), BL2 at 10 μM showed increased inhibition compared to BL3 and BL4 (10 μM). The BL2 (structure shown below) fits the independent and dependent DNAi motif claims. Both BL3 and BL4 contained a single mismatched base meaning neither sequence had 100% homology to its complementary strand. This demonstrates that many times even a single mismatch to the complementary strand decreases the inhibitory effects of a DNAi oligonucleotide. The mismatches for BL3 and BL4 are noted below with the mismatched letter highlighted and bolded. It should also be noted that a 20-mer version of BL2 demonstrated similar significant inhibition (data not shown) as the 24-mer version of BL2 shown in FIGS. 50, 51, and 52.

BL3: ACCGGCGCTCGGCGCGCGGA (SEQ ID NO: 13825)(needed to have a G in place of the C for 100% homology)

BL4: GACGCGCCGGGCCGGGCGGA (SEQ ID NO: 13826) (needed to have an A in place of the C for 100% homology)

In FIG. 52, as a counter screen to test for nonspecific toxicity, BL2 and BL7 were tested at 10 μM in NMuMG (a normal murine mouse mammary gland cell line) and measured at 24 and 96 hours post exposure. As would be expected, BL2 has no cytotoxicity against a normal, nontumorigenic mouse cell line because it was designed for homology with the human genome and only has a maximum of 67% homology across the entire mouse genome. BL7, however, has approximately 90% homology across the entire mouse genome. This demonstrates that duplication and high overlap with non-targeted regions of the genome leads to non-specific cytotoxicity.

The secondary structure for BL2 are shown in FIG. 53. Sequence 302 (BL2) is shown in FIG. 53.

In FIG. 54, In HCT-116 (human colorectal carcinoma), BL9 produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The BCL2 sequence BL9 will not form a secondary structure under physiological conditions.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13683) CCTCCCAAAGTGCTGAGATTACAGGCATGAGCAACCACACCTGGCCGATA CATACCTATATTAAACATTAGTATGTTCATGTTAGAATAATGTACCTTTT GAATTTCATAAACTTGGAGAATATTTATATTGATGATGGATGAAAGAACT TTCTTGGATGGATGAAGAGAGTAACGCTGTGAAACAACCAGCAGGTGGCG AAAACTGGCAATCAAAAGCTTTTTGTTTGGTGGCCTGGGGAATGAAGACG GAAAGAAAACACAGGCCATTCAGACTCTTGATACAATCTCCATTCCCTGC ATCTTGTTTTTTTTCTCTTCCTGGTGCCACGCTACTTGTAGAATCCAACC AGGTAAAGCTGCCAAAAGGGTCATCCATTGGCTCTTACAAGTAGAAAACA TCTTGGAAAGTGAAAAGTCCACTGTGCATATGTTTGTAAGGTTGTTGGAA GGTCTCAGGCATAGATCTAGGATTCAAATCCAGTTACTCCTGCCTCAGGC TGGTGTTCCCTCCCCCACCTCAGCATTGCCCAAAGACGAGTAGTCAATGT CACATACTTCCTGGGAATACTTGCCCTTATGCTTTAAAATGGAATCTAAT AAACATGGAATCTGAACGCAGGAATGGTTTCACTCTTTCATTTGAAAGAG TATCTAGAACATTCCCAGGGAAAATATAACCCCTAGCCAAAGACTGCAAT ACAGACCTGTCTCAAGACTGATTATAGCCAAGATGCCACATAAGGAATCA GTCTGGGAAAATCCATAGAGTGAGGCTCTGTGGGAGCAAAGGAAGACGAA AATCAGTCAGCTTTTCTTTCTCTGGAAGTAGGGGATCCGTTTCCTTCTGG CTGCCCCCTTTGCAGAAGTACAGTTTCTTTTGCAGGTTTGTCCTATCATT TCCTCACTCATATGCTGAGTATTAGGAGCTTGAAGCCTTTCAATTCCTCT TAGGTAATTTTGGGGCTTTAAAATACGCTTTCAAGATTTCTAAACCATAC TGTTGTGCAATTGGTATGAATTTATGTGAGAACATTTATTCTAGGTCAAT CTATACCCAGTGTCTATCCAGACCAAAACACCTCCCACGCGCATAAAAGG GACTCTGTCCCAACCATCAGAAGGGCAAGAAGGAGGATCTCCTTTCATCC CCTCTTGCCTGGATAAGAAATTTGTACCCAGGCCCCCATTCCTATGTGAG AGAAGTTGGCTTGTTGGGCTGATGGGATACAATAAATGAAGAAATAAAAT AAAAACACCCAAGAGAGATGGCAGTGCGTATAGTCCCAGCTATTCATGAG GCTGAGGTGGGAGAATCCTTCGAGCCCAGAAGTTCGAGTCCAGCCTGGGC AACATAGCAAAAGCCATCTCTTAAAAAAAAAAAAAAAAGGCCAACTAAGT AAAAATTAAAAAAATCATAATTTGGTGTGCTTTTCTGGCTTTTTAAAGAA TGTTTTGATTTTAGAGTAGGAATGAGACAAAATAAAGATGTCAGGCAGGG CACAGTGGCTCATGTCTGTAATCCCAGCACTTTGGGAGGCTGAGGCAGGC GGATAACGAGGTCAGGAGTTCGAGACCAGCCTGGCCAGTATGGCGAAACC CCGTCTCTACTAAAAATACAAAAATTAGCAGAGCGTAGTGGCGTGCACGT GTAATCCCAGCTACTCAGGAGGCTGAGGCAGAATTGCTTGAGCCTGGGAG GCAGAGGTTGCAGTGAGCTGAGATCGCACCACTGCACACCAGCCTCCAAG ATACAACAGAGCAAGACTCTGTCTCAAAAAAAAAAAAAAAGTCATAGCAT ATTTGTACACATTGTAGTACTCATTTGTCATCTTTCTTGACCCCAATAAT CCAGTGTCCCTATATATTTGCACTCGAGCCCTATTAAGTAAGCCGCTGTG CTTCTAGAAGACCTTTTTCTTTTCTTGGTGCTTTGTCAAAGACTCTTGGA GATAAAAATACACACGTGCAACTTGTTTGTCCTCTTGTCCTTTTTTGCTA GGGGCTATTCATGCTGATTAATTTAAAACTGTCTGCTTGCGCGTACACAC GTCTGCGAGTGTGAATGTGTATGTGTGTATCTATGTACCTCATTTGAGAA AGTGCGGCCAACTAGGATTGGCTACGAGGCAAAGGTGGAGACCTTTAGGA GCCCACCCACCCCAGCGTTAGGACGGTGGGCCTGAAAGTTACTATATGGA AGTCCTCATCGTGTAGCACTAAACCAGTGTAAAAGGTGTTAGGGACAGAG GGAAAACATTGACTTAAACTGTCGTAAAGCCCTTGATAAACCCCTTCCCT GGAGCGCTGAGTTCTGCATGGCCTGGGCCACGGACTAGGTGTTCAGGTGG ACACGGGCGGGGATGCGCGTGCGTGTGTAGTGCGCGGACACCTAGGAAGC TACTTGAAAGTAAACACCACGCTCGGGGCGTCCCTAGACATTGCTTAAAA CGTGCAGAGTCACCTGTCTTCACAGCAGGGCAGCGCTGAGGTCCCACTGC TGGGGGCGGTGGGGGGCGGCATTGGCCTGGGTCTTCCCCCGGCGGCCGAG CGCCGGTAACACAACGTGTGTGTGTGTAGGCGCGTGTACACACTCTCATA CACGGCTAGAAAGGGTCCAGGCGACACACACACTCCCACATACACGGCTA GAAAAGGTCCAGGCGAGACACACACACACACACACACACACACACACACT CCACACACACTCACACGGCCAGAAAGGGTCCAGGCGGTTCCCGGCGCTTT TCCAGCCCTTGTTTTCATGGCGCACCCTCCCGCCAGCCGCCCCCCTCCGC ACTCCGTCGTCCGCCCGGCCCGGCCGCGTGCGGTTCCCCGGGAGCCCCCA CCCCGTCGCGGACCCCAGCGACCACCAAGTCCGCACGCGGCCTGCCGCAG GCCTGAGCAGAAGGCCCCGCGCACACCCACCGCGCCGCGGCCGCGCGGGA GGCCTGTGCCGCCCGCGCCACCCACTGGCCGGGCCCCGCGGGCGCAGCGG AGCGGGCGGGTGGCCGGCCCGGACGCGCCCTCCCCGGCCGCGGCCCCGCG CGCCATGTGCCCCCGGCGGGACGCGCCACTCCCGGGCCTGCCGCGGCGCC TTTAACCCGGGCCAGGGAGCGGGGCGGAGGGGGCGGTCGGGTGGCTCAGA GGAGGGCTCTTTCTTTCTTCTTTTTTTGAATGAACCGTGTGACGTTACGC ACAGGAAACCGGTCGGGCTGTGCAGAGAATGAAGTAAGAGGACAGGCACC ACAGCCCCGCTCCCGCCCCCTTCCTCCCGCGCCCGCCCCTCCGCGCCGCC TGCCCGCCCGCCCGCCGCGCTCCCGCCCGCCGCTCTCCGTGGCCCCGCCG CGCTGCCGCCGCCGCCGCTGCCAGCGAAGGTGCCGGGGCTCCGGGCCCTC CCTGCCGGCGGCCGTCAGCGCTCGGAGCGGGCTGCGCGGCGGGAGCTCCG GGAGGCGGCCGTAGCCAGCGCCGCCGCGCAGGACCAGGAGGAGGAGAAAG GGTGCGCAGCCCGGAGGCGGGGTGCGCCGGTGGGGTGCAGCGGAAGAGGG GGTCCAGGGGGGAGAACTTCGTAGCAGTCATCCTTTTTAGGAAAAGAGGG AAAAAATAAAACCCTCCCCCACCACCTCCTTCTCCCCACCCCTCGCCGCA CCACACACAGCGCGGGCTTCTAGCGCTCGGCACCGGCGGGCCAGGCGCGT CCTGCCTTCATTTATCCAGCAGCTTTTCGGAAAATGCATTTGCTGTTCGG AGTTTAATCAGAAGAGGATTCCTGCCTCCGTCCCCGGCTCCTTCATCGTC CCCTCTCCCCTGTCTCTCTCCTGGGGAGGCGTGAAGCGGTCCCGTGGATA GAGATTCATGCCTGTGCCCGCGCGTGTGTGCGCGCGTGTAAATTGCCGAG AAGGGGAAAACATCACAGGACTTCTGCGAATACCGGACTGAAAATTGTAA TTCATCTGCCGCCGCCGCTGCCTTTTTTTTTTCTCGAGCTCTTGAGATCT CCGGTTGGGATTCCTGCGGATTGACATTTCTGTGAAGCAGAAGTCTGGGA ATCGATCTGGAAATCCTCCTAATTTTTACTCCCTCTCCCCGCGACTCCTG ATTCATTGGGAAGTTTCAAATCAGCTATAACTGGAGAGTGCTGAAGATTG ATGGGATCGTTGCCTTATGCATTTGTTTTGGTTTTACAAAAAGGAAACTT GACAGAGGATCATGCTGTACTTAAAAAATACAAGTAAGTTCTCTGCACAG GAAATTGGTTTAATGTAACTTTCAATGGAAACCTTTGAGATTTTTTACTT AAAGTGCATTCGAGTAAATTTAATTTCCAGGCAGCTTAATACATTCTTTT TAGCCGTGTTACTTGTAGTGTGTATGCCCTGCTTTCACTCAGTGTGTACA GGGAAACGCACCTGATTTTTTACTTATTAGTTTGTTTTTTCTTTAACCTT TCAGCATCACAGAGGAAGTAGACTGATATTAACAATACTTACTAATAATA ACGTGCCTCATGAAATAAAGATCCGAAAGGAATTGGAATAAAAATTTCCT GCATCTCATGCCAAGGGGGAAACACCAGAATCAAGTGTTCCGCGTGATTG AAGACACCCCCTCGTCCAAGAATGCAAAGCACATCCAATAAAATAGCTGG ATTATAACTCCTCTTCTTTCTCTGGGGGCCGTGGGGTGGGAGCTGGGGCG AGAGGTGCCGTTGGCCCCCGTTGCTTTTCCTCTGGGAAGG ATG

BCL2

Apoptosis also plays a very active role in regulating the immune system. When functional, apoptosis causes immune unresponsiveness to self-antigens via both central and peripheral tolerance. When defective, it may contribute to autoimmune diseases (Li et al., Clin. Dev. Immunol. 13 (2-4): 273-82 and reviewed by Tischner et al., Cell Death and Disease (2010) 1, e48), such as type 1 diabetes, manifested as aberrant T cell AICD and defective peripheral tolerance. Dendritic cells are the most important antigen presenting cells of the immune system such that their activity must be tightly regulated by such mechanisms as apoptosis and their lifespan may be controlled in part by BCL-2. Other inflammatory diseases include inflammatory bowel disease, psoriatic arthritis, lupus, heart disease, and Alzheimer's and schizophrenia.

Given its biological importance, BCL2 is a prime candidate for targeted therapies. Numerous approaches that block or modulate production of BCL2 at the DNA level (e.g., retinoids and histone deacetylase inhibitors), RNA level (targeted antisense oligonucleotides such oblimersen and SPC2996 or siRNA approaches), or the protein level (gossypol, obatoclax, ABT-737, ABT-263, ABT-199) have been reported and a few have entered clinical development.

30. CMYC. Myc (c-Myc) is a regulator gene that codes for protein that is a transcription factor. In the human genome, Myc is located on chromosome 8 and is believed to regulate expression of 15% of all genes (Gearhart et al., N Engl J Med 2007; 357:1469-1472). CMYC activates expression of many genes through binding on consensus sequences (Enhancer Box sequences (E-boxes)) and recruiting histone acetyltransferases (HATs). This means that CMYC is activated upon various mitogenic signals such as Wnt, Shh and EGF (via the MAPK/ERK pathway). By modifying the expression of its target genes, Myc activation results in numerous biological effects. CMYC has the capability to drive cell proliferation (upregulates cyclins, downregulates p21), but it also plays a very important role in regulating cell growth (upregulates ribosomal RNA and proteins), apoptosis (downregulates Bcl-2), differentiation and stem cell self-renewal. CMYC is a very strong proto-oncogene and it is very often found to be upregulated in many types of cancers. Myc overexpression stimulates gene amplification (Denis et al., Oncogene 6 (8): 1453-7), presumably through DNA over-replication.

It can also act as a transcriptional repressor. By binding Miz-1 transcription factor and displacing the p300 co-activator, it inhibits expression of Miz-1 target genes. In addition, myc has a direct role in the control of DNA replication (Dominguez-Sola et al., Nature 448 (7152): 445-51).

Mutated CMYC is found in many cancers, causing it to be constitutively expressed thereby driving the unregulated expression of many genes involved in cell proliferation. A common human translocation involving CMYC is t(8; 14) which is critical to the development of most cases of Burkitt's Lymphoma. Malfunctions in Myc have also been found in carcinoma of the cervix, colon, breast, lung and stomach (Prochownik, 2004; Expert Rev Anticancer Ther.; 4(2):289-302).

Because CMYC is part of a dynamic network whose members interact selectively with one another and with various transcriptional coregulators and histone-modifying enzymes, it is an attractive therapeutic target. Several approaches including small molecules, peptides, and oligonucleotide therapeutics have been pursued. However, knowledge of which pathway should be attacked (c-Myc transcription, translation, interaction with other myc network members, DNA binding and transcriptional activation) is crucial. Clinical efficacy will likely require intervention at several levels, perhaps in combination with traditional chemotherapeutic drugs or agents that target other oncoproteins (reviewed by Levens, 2010; Genes and Cancer 1: 547).

CMYC

Protein: CMYC Gene: CMYC (Homo sapiens, chromosome 8, 128748315-128753680 [NCBI Reference Sequence: NC_(—)000008.10]; start site location: 128748840; strand: positive)

Gene Identification GeneID 4609 HGNC 7553 MIM 190080

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 7551 CGATGAGGGTATTAACTCTGGC 335580 7552 CGGGGGTCCTCAGCCGTCCAGACC 518 7602 CGCTTATGGGGAGGGTGGGGAGGG 634 7603 CGGTGGGCGGAGATTAGCGAGAGA 559 7606 GGCGCTTATGGGGAGGGTGGGGAGGG 632 506 CCTGGCACGTGTCCCTGGTCAAG 3482 507 CACGTGCGGCCTGTCAAGAGATGA 5926

Target Shift Sequences Relative upstream Sequence location to gene ID No: Sequence (5′-3′) start site 7551 CGATGAGGGTATTAACTCTGGC 335580 7552 CGGGGGTCCTCAGCCGTCCAGACC 518 7553 GGGGGTCCTCAGCCGTCCAG 519 7554 GGGGTCCTCAGCCGTCCAGA 520 7555 GGGTCCTCAGCCGTCCAGAC 521 7556 GGTCCTCAGCCGTCCAGACC 522 7557 GTCCTCAGCCGTCCAGACCC 523 7558 TCCTCAGCCGTCCAGACCCT 524 7559 CCTCAGCCGTCCAGACCCTC 525 7560 CTCAGCCGTCCAGACCCTCG 526 7561 TCAGCCGTCCAGACCCTCGC 527 7562 CAGCCGTCCAGACCCTCGCA 528 7563 AGCCGTCCAGACCCTCGCAT 529 7564 GCCGTCCAGACCCTCGCATT 530 7565 CCGTCCAGACCCTCGCATTA 531 7566 CGTCCAGACCCTCGCATTAT 532 7567 GTCCAGACCCTCGCATTATA 533 7568 TCCAGACCCTCGCATTATAA 534 7569 CCAGACCCTCGCATTATAAA 535 7570 CAGACCCTCGCATTATAAAG 536 7571 AGACCCTCGCATTATAAAGG 537 7572 GACCCTCGCATTATAAAGGG 538 7573 ACCCTCGCATTATAAAGGGC 539 7574 CCCTCGCATTATAAAGGGCC 540 7575 CCTCGCATTATAAAGGGCCG 541 7576 CTCGCATTATAAAGGGCCGG 542 7577 TCGCATTATAAAGGGCCGGT 543 7578 CGCATTATAAAGGGCCGGTG 544 7579 GCATTATAAAGGGCCGGTGG 545 7580 CATTATAAAGGGCCGGTGGG 546 7581 ATTATAAAGGGCCGGTGGGC 547 7582 TTATAAAGGGCCGGTGGGCG 548 7583 TCGGGGGTCCTCAGCCGTCC 517 7584 CTCGGGGGTCCTCAGCCGTC 516 7585 GCTCGGGGGTCCTCAGCCGT 515 7586 AGCTCGGGGGTCCTCAGCCG 514 7587 CAGCTCGGGGGTCCTCAGCC 513 7588 ACAGCTCGGGGGTCCTCAGC 512 7589 CACAGCTCGGGGGTCCTCAG 511 7590 GCACAGCTCGGGGGTCCTCA 510 7591 AGCACAGCTCGGGGGTCCTC 509 7592 CAGCACAGCTCGGGGGTCCT 508 7593 GCAGCACAGCTCGGGGGTCC 507 7594 AGCAGCACAGCTCGGGGGTC 506 7595 GAGCAGCACAGCTCGGGGGT 505 7596 CGAGCAGCACAGCTCGGGGG 504 7597 GCGAGCAGCACAGCTCGGGG 503 7598 CGCGAGCAGCACAGCTCGGG 502 7599 CCGCGAGCAGCACAGCTCGG 501 7600 GCCGCGAGCAGCACAGCTCG 500 7601 GGCCGCGAGCAGCACAGCTC 499 7602 CGCTTATGGGGAGGGTGGGGAGGG 634 7603 CGGTGGGCGGAGATTAGCGAGAGA 559 7604 CCGGTGGGCGGAGATTAGCG 558 7605 GCCGGTGGGCGGAGATTAGC 557 7606 GGCGCTTATGGGGAGGGTGGGGAGGG 632 13684 CCTGGCACGTGTCCCTGGTCAAG 3479 13685 CTGGCACGTGTCCCTGGTCA 3480 13686 TGGCACGTGTCCCTGGTCAA 3481 13687 GGCACGTGTCCCTGGTCAAG 3482 13688 GCACGTGTCCCTGGTCAAGT 3483 13689 CACGTGTCCCTGGTCAAGTA 3484 13690 ACGTGTCCCTGGTCAAGTAG 3485 13691 CGTGTCCCTGGTCAAGTAGG 3486 13692 ACCTGGCACGTGTCCCTGGT 3478 13693 TACCTGGCACGTGTCCCTGG 3477 13694 TTACCTGGCACGTGTCCCTG 3476 13695 TTTACCTGGCACGTGTCCCT 3475 13696 ATTTACCTGGCACGTGTCCC 3474 13697 AATTTACCTGGCACGTGTCC 3473 13698 AAATTTACCTGGCACGTGTC 3472 13699 GAAATTTACCTGGCACGTGT 3471 13700 GGAAATTTACCTGGCACGTG 3470 13701 AGGAAATTTACCTGGCACGT 3469 13702 AAGGAAATTTACCTGGCACG 3468 13703 CACGTGCGGCCTGTCAAGAGATGA 5928 13704 ACGTGCGGCCTGTCAAGAGA 5929 13705 CGTGCGGCCTGTCAAGAGAT 5930 13706 GTGCGGCCTGTCAAGAGATG 5931 13707 TGCGGCCTGTCAAGAGATGA 5932 13708 GCGGCCTGTCAAGAGATGAG 5933 13709 CGGCCTGTCAAGAGATGAGG 5934 13710 TCACGTGCGGCCTGTCAAGA 5927 13711 GTCACGTGCGGCCTGTCAAG 5926 13712 AGTCACGTGCGGCCTGTCAA 5925 13713 AAGTCACGTGCGGCCTGTCA 5924 13714 CAAGTCACGTGCGGCCTGTC 5923 13715 TCAAGTCACGTGCGGCCTGT 5922 13716 TTCAAGTCACGTGCGGCCTG 5921 13717 CTTCAAGTCACGTGCGGCCT 5920 13718 CCTTCAAGTCACGTGCGGCC 5919 13719 TCCTTCAAGTCACGTGCGGC 5918 13720 TTCCTTCAAGTCACGTGCGG 5917 13721 ATTCCTTCAAGTCACGTGCG 5916 13722 AATTCCTTCAAGTCACGTGC 5915

Hot Zones (Relative upstream location to gene start site)   1-1880 2150-2240 2420-3050 3230-4130 4310-4400 5900-6000 335000-336000

Examples

In FIG. 55, CM7 at 10 μM showed statistically significant inhibition compared to control values in MCF-7 (human breast cancer cell line). CM7 (structure shown below) fits the independent and dependent DNAi motif claims

In FIG. 56, CM7 at 10 μM showed statistically significant inhibition compared to control values in MDA-MB-231 (human breast cancer cell line). CM7 (structure shown below) fits the independent and dependent DNAi motif claims.

In FIG. 57, In MCF7 (human mammary breast cell line), CM7, CM12, CM13, and CM14 produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated control values. The CMYC sequences CM7, CM12, CM13, and CM14 fit the independent and dependent DNAi motif claims.

The secondary structure for CM7 is shown in FIG. 58. Sequence 317 (CM7) is shown in FIG. 58. The secondary structures for CM12, CM13, and CM14 are shown in FIG. 59, FIG. 60, FIG. 61.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13723) TGATTGTGGCCAGGCACTACAGCTCACACCTACAATCCCAGCTACTCTGG AGGCTGAGGTGCGAGGATGGCTTGAGCCCAGGAGTTCAAGACCAGCCTAG GCAACATAGTGAGACCCTGTCTCTAAAAGGTTTTCTAAAATTAGCCAGGT GCATATGCCTGCAGTTCCAGATTCTCAGAAGCCAGAAGTGGGGAGGATCT CTGGAGTTCAGGAGTTTGGGACCACGGTAAGCTATGATTGTGTTACTGCA CACCAGTTTGGGTGACAGAGCGAGACCCCTTCTCTCAAAACAAATAAATA AGATTGTGGTGATAGCTATACAACCCTGTGAATAACTAAAAATTGTTGAA CCATGCACTTTAAGTGCATGAATTTTATGGCATGTGAACTTTATCTCAAT AAGGCTGCCATTACAAAGCTAAAAAGGGAGGCAGGTGCATGAGCACATAT ATGTCTAATATTAGCTAAAATAGTATCACCATTATTAAATAAACTTTTAA AAAATACCTTCTTTCTGAGAATGCAATTCTTCCTTATAATCAGAACCATG AATATACCAGGAAACTTTTTAAATCAGGGAACAAATGCCTACGAAGGACA GGCACAAGCCAGAAAGGGACTATGGATGAATTAAGTGGGCTGAGCATATG GGAGCGGTGGAGACTGGGGCAAACTGAACAGCTCCTGGCCCTTTTAAAAG AAATCGGCTGCTCCTCAACTTCCATCCACTTCTGAATGCAGTTCCAGAAT TACCAAATCTGCCTGTTTAAGGAAAGGCACAAATTCAGATTGTTAATGTG AAATCTATTGACTTGTAAGTGTTGGCACCTATTTTTAAATGTTATAAATG CTGAGAGGGTCAAAACCTGTCATCCAAGCCAACCTACCAGTAAGCAAGAC TTGGTCCTCAAGCAAGTTTGCTGCCTCTGCTTTGAGTACTTTAGCATGAC TTTCAAAACCTCCCACCTCCCCCTCGCCCTGCCTAAACCCACTTTACCCC TCACCACCACTGCAAGAAGTTATCCAAGCTATGAAGAGAGACAGAAGAAT TCATACATAAATAAAGAGTCCCAAAACATTCTCAAAGATGCCAAAGTCAG GCTAGGGTGGCATGGAGAGGGAGTGGGGCATAAAGTTTTTGATTCCTAAT CTAATTAGAGAGCCCTATAACAGATTCTTTGTTCAAAGACCAAATTTAAT TTACAATTTTATATCTCCAGTGAAGTCAGCTTTTATTAATTTCCAGCACA ATATTTGGATATACTGGCCAGAACTTCAATGAGTTCCTATTTAGTGTTTA ATCTTCTAATGCATTCCAATTAATTATTTCAGTTTTATGGCAAACTGTCT TCAGCCAACATCCAAGCTGGACACCCCATGCCTCTCCACTCACCCAAAAA ACCAGCTCGGGAGGTGTCAATATAATGACTTAAGATGCTGAATGGTAAAG GACAGGATTGGAAGGAAATTGCGCCTGCAATTATGCACTAATGCTTCACC AGAGAAGCAGATGGCATTCCTTGCATAAATTATTATTTATCCTTGGAATT CCCCTCTGCCTATTACCAAATCAACCCTTGAAAACAAGTCTTTGTTGGGT CTGTGAAGTCCCCTGGCCAGTTTCCAATGTCTGCTCCCTCCCTCACATCC CACCCTCCAGAGCTGCAGCGAGGGTAAGAACTCCAACATGGCCCACAGGC AAGGGTTTCCGAAAGCATCGACGTTCTAAATACATTTGGACGGAGGTGCA CAGAAAGGAGTCCGCTTTATTTTGCAGACTGGGAATCCAGATGCAATGAC CACAGGCAGAAAGCATGGAGCAGAACCTCCCAGCCTCGGCTGTACCCCCA GTGATAAGGCTTGCCACGTGTGGACGTCACCAGGTTGCCCACCACAGCAC GGGGCTTAGGCTGTACTGTGCATTCTCTCATGGAATCCTTGAACAAGGAT TGAGGTGGGCAATGATGTTCCACTTTGAGGAAATGAAATGAAGAAACCAG AGACTCTGAGACAAAGAAAAGGGCTTTGGGTTTTTTTGTGTTTTTTGGCT TTTTATTTATTTATTTATTTTGTACAGATGAGGTCTCACTTTGTTGCCCA GATTGGTCTCAAAGAATGGTGCTTTGGATTAGATCTTATTGTGATGAAAA ATAAAAAAAAATTAAAAATTTTTTAATTTAAAAAGAATACTGCTTTTTTT TTTTTTTTTTAACAGGGTCTCTCTCTATAGCCCCTCTCTATAGAGTGTAC AGTGGCACAATCTCAGCCCGCTGCAACCTCTGCCTCCCAGGTTCAAGCGA TCTTCCCACCTCAGCCTCCTGAATAGTTAGGACTACAGGCATGTGCCACC ACGCCTGGTTAATCTTTTGTAGAGATGGGGTTTCGCCATGTTGCCCAGGC TGGTCTTGAACTCCTGAGCTCAAGCGATCTGGCCACCTCAGCCTCCCAGA GTGCTGGGATTACAGGTGTGAACCACCATGCCCAGCCAGAACACTGTTAA CCTTAACATCAACAGGCAGCTACCATTTTCGAGTGCCTGCAATGTCATTT AACCTTTAGGAACAGCTCTGGGAGACAGCTATAGTTGTTGCCATTTTCTG CAGATTGAGAAACTGAGGCTCAGTTAAGTGACGTAATTCTAAGGCACCAC ACCCAGTCAAGCGCAGTGACAGAATTCGAACTCTGGCTTGTAGGGATTCA CAGGACTGCCAAAGCTTACGCTAACCCATTTCTTCTCCTGTGCACCATCA TTGCCTCATTCTCTGCCCTCATTTTCTTTATTTATTTTTATTTATTTATT TTTCTTTTTTTGAGATGGAGCTTCACTCTTGTTGCCCAGGCTGGAGTGCA ATGGCACGATCTCGGCTCACTGCAACCTCCACCTCCCGCGTTCAAGAGAT TCTCCTGCCTCAGCCTCCTGAGTAGCTGGGATTACAGGCATGCACCACCA CGCCCAGCTAATTTTGTATTTTTAGTAGAGACGGGGTTTCTCCATGTTGG TCAGGCTGGTCTCGAATTCCCTACCTCAGGTGATCCACCCGCCTCGGCCT CCCAAAGTGCTGTGATTGCAGGCGTGAGCCACCGTGCCCAGCCGCTCTGC CCTCATTTTCTCCCCAAAACCAAAGTCTACTTTACAAGCACAGATATTAC TAACTTGTCTTACGAAACTTTCCAGAAGAAAGAGAAAGAATATATGTTTT ACCAAGCCCCTTGGAGGACAAGGATTTGTTTCTGTATCCACTGTCTCGAT ACTCATGGTGCCTTTTACCCCTTGGCATTATGCCCCAGGAAAGTGGCAAA AGTAAGAGGTAACCTCTCCTTCCTTCCTTATTTCCCTAAGGAAATTTGCT CTGGTCACCAGCAGCAGAGAAATAGAAAGCGCCGGGCACCTGGCTCGACT GGGGCAGTGACAGGGCAGAGGCGGCCCAGGTTATGGTATCAAAAGGTTTC TGGTGCTGAATCTCATGACTACTATTCACCGTGTGAGTTTAAGCAAGTCC CTGCAACACCTCAATTTTCCCCATCTGTTAAATGGAATTTTAACCTACAC CTCCTAGGATTACTATGGAGATTTAAGGAGGCAATGCAGTGGGGCTTTCT AACCTTTTTAACTCACTGAGATACATTTCCCGTATCGTCCAAGTGCAGAC ACACACACACACACACACACAGAGAGAGAGAGAGAGAGAAAAGAATACTT CATCTGCAACACACTTTGATATTTTCTGTGCCAGCCCATTTTGTGAAATT GCTCATCATGATTCATTAAATTCATTTCTTATTTACTATTTTTAAATTTT TATACATGCAGGGGGCTCAAGTGAGGATTTCTTTCATGTATACATTGCAT AGTCGTCAAGTCTGGTCATTAAATGTATTCATTATCCAAATAGTGAACAT TGTTAAATTGATTTCATGATCCACTAAGGGGTCATCATTTGCCATTTTAA AACTCTGACAGTATGAGCTTCTCCCTAGCCCAGTTCCTGTTACCATCTTC CCATTCTTCCCTTCCTTCTTCAATTCAGATAGGATTTTCCTCCAGAGGGA TTATAAAGTTGCGAGGAAAGCGCCTGCAGGGGGTGCTGTTCCACACTGTT GTTGAAGTGTGGTTTGGTTTTTATTTCGTTGCATTTGCTTTTCGGTCAAT GAGGGCAATTCATCTGGAATGACCCCCATCCTCGTCACCCTTGCTCCAAC GATGTTGGGGCCCAGCTCATCAACAAGGACACCTGAACAGAGCCCTACCC ATTGATGGAACCGAAGCAAGGGCAAGGAAGAGTTCTCAACCCTTCTCTCT ATATACGATTAAAACTGGGTTAGGCTAGGTGTGCCCTCAGCTCAGAAGCT CTCTCTAATAGCATTCCTTCACTAAGCACTTACAGAGTGCCTACCACGTG CCAGGCATTGTGCTGGGCTCTGGAGACCACCTACTCTGTGAATGGCACCT TGAGGCTTGATGGGTGAGAACGCGAGTAAAACACAATCCATACTGACCCC AGAAGCTTCTCCTCAAGGAATCAGACATTAAAAAGCACAAAAACTATAAA GTTGATTTTTTTTTTTTTTTTTTTTTTGAGACAAGAGTCTTGCTCTGTCA CCCAGGCTGGAGTGCTGTGGCACCATCTGGGCTCACGGCAACCTCCACCA CCCAGGTTCAAGCAATTCTCCTGCCTCAGCCTCTCGAGTAGCTGGGATCA CAGGCATGCGCCACCATGCCCCGCTAATTTTTGTCATTTTTAGTAGAGAC AGGGTTTCACCATCTTGGCCAGACTGGTCTCGAAATTCTGACCTCGGGTG ATCTGCTCACCTCAGCCTCCCAAAGTGATGGGATTACAGGCATGAGCCGC TGCATCTCTGGCCAAAACTTGAATGTTTGTTTGTTTTGAGACAGGATCTC ACTCTGTCATCCAGACTGGAGCACAGTGACACAATCTTGGCTCACTGCAG CCTCAACAGCCACGGCTCAAGCAATCTTCCTCCTCCACCTCAGTTTTCCA AGTAGATAGGATTACAGCCATGAGGCACTGCACCCAGCTAATTTTTTTTT TTTAATTTTTTTGTAGAGACAGGGTCTTACTGTGTTGCTCAGGCTGGTCT CAAACTCCTGGGCTCAAGTGATCTGCCGGCCTTGGCCTCCCGAAATGCTG GGATTACAGGTACGAGCCACCACGCCTGGCCAAACTTGTATTTTCTAAGA CAGAAGAATGAGGGGATGGTTTAAACTCTCAAGGGAAGGGGAAAGGATCA TGAAAAGCTCCTACAGGAAGATGCTTGAGTTGGATTACTAAGACATATGA GCAGAGATGGCAGGCTGGCAGCCTGAGGGCCACCTCTGCCCATAGACATG CTTTGCTTCTCCATATCATTTTTTTTCCCAACACACTGCTGCTGGCTTGA AATCTCCATATAATTCTTACAATAAGTTGTTAACATTTTAAAACCTGGAT TTCCACCTTCCCTGAAAAACTGGAAGCATTTCCACCCATGGGCCCATATT TCAGGGTAACCACCAGAGCAGGTGCCAAATGGGAGCCACCAGACCTACAC AGGCAAATGCTCTCCAGTTTACCAGTCTCCACCACTCCCTATTGTATTCT TCGTTTACATTTCCTGCCAAACCTCTGTAAGCATCTGAGTTGGCAACCCT TGATGTGTTAGCGGAAAATGTGGATCAGAAGTTAGAAAGAGTTTCTAAAC CTGGTTGTTGATTTACGCTTTATGCTTTGAAGGAAAACAGTTTTTCCAAT GCCCAGATCCACTCACCAAGACAAAAAAAAAAGCAAGCTGTAGATTTCAG TAGCAGCCTTGTCTAGCCAGCAATAAAGGTGCCCTGGGTTTCCAGGACCA CACCCCAGGGATTAGCCCCGGGGCATCATATGAATTCAGTGAAAGGCGGG AAATCCTAACATAAAGCGTTGATTCGTATTAAATAGGAACAATGCCTAAT TCTGCCTTCCTGAACTTCCAGAATTTTGCTTTTTCTGAATAGAGTGATCT GCAAAACAGCATACACTTGGAATAGTAAGTCGTGCAAGAGTTGGAGACAG GAAGGGGGTGGGTTTGGAATTGTCTCCAAACATTAGATAATCTCTTTGTG ATTCTAAACCTCAACTTGACAAGCTTGTATTAGTCCACAATTTTTCACAC TTGATGAAGTGATAAAGGACATCAATTTCATGGAACTCACTATGAAACAC CATGCAATATTGATACATTTAACTTAAAACAGCTCAATACATAACTTTCT GCTAAATCTGGAACTCACATTAACAATTGCTAACATTTGCTGAGTGTGGG CCAGACAGCAGGCTCTGTGCTGAATGCCTTATCTCACTTAATTCCTGTAA CACCTTCAATAAGATAGGTGCTACAATTATAGTAATCCCATTTTACAGAT GAGAAAAGTGAGATTCAGAGAGGTCATGTGACTTGACAGATTTATCAGGT GATCATGACAGAGTAGTCCTCCAACCAAGCTGATTCAGCAACCCGTCCTT ATATTCTAGATTCTTGTGTAGCCAAAAAGTTATTGAGAAAGTCTGCCCAT TGACTTCATTCTCTTACCCAGTGTAGAGTCAGCATACATTCATTCACATT AACTATGGGCCAGACTTGATTCCTGGCCTTGGGACTTTTTTTTTTTTTTG GCAGGGGCTGATAACATTCTATTTTATTTATTTATTTATTTCTCTCTCTT TTCTTTTAATTATACTTTAAGTTCTGGGATACATGTGCAGAACATGCAGG TTTGTTACATAGGTATACACATGCCATGGTGGTTTGCTGCACCTACCAAC CCATCATCTACATTAGATATCTCTTCTAATGCTACCCCTCCTCTAGACCC CCGGGACATTTATAATCTCATGAAGAAGAGAAAAAGGAGGCCTTTCTCTG ACAGCTAGAAAACCACAGTTAGTCTATTTTAGCCGGAGACCCTGGATTCT ACCCTGAGAACAAAGGTTTATGTTTCAGCAGCTTAATTAGAGGTTTTCCA GAACTTTTTCTGGCTCCATGCTTTTATGATTCTGTAAGATGATCATGGGA AAAGGAAGAGTCCACAGAGAAAATGGGGCTTGAACTTGGGCTGGGAGGAA AGGTGGTTCTTAGATAAATCAAGAAGAGAAGAGACAGTAAGTCTGGGGAA CTGCCTGAACCAAAGTGCTGAGGTGGAAACTTGTGTGTCACTCAGAGTGG CTGTAAATAGACCTGTTTCTCTGAAGTGCAGAGTTGGTAAGAAATAGGGT AAGATAAGGAGGAGGCCAGATGCATGAGGGCTTGGAATTCCAAGCTCATA ATGGAGAACCTCATTTTGGACCATGGGGGTCAACTGAAGAATTTTAAATG AAGAGGAAAATTAATCAGTGTGCAAGGTTAAATGGAGTGGCAGAGACTAG GAGCTATTAGGAATCTACTGCAAGATGATTCTAACAGCCATAGGTAGTGG GTAAAAGAGGAAAGTGAGCCAATAAGGGAAACAGAAGAACAAGTTGAATA TGTGGGAATATAATCAGGAGAATGTGGAGTGAACCCAGGGATTCTCAACC TCAGCACTGGTGACATTTTGAGCCCCACTCTCTGTTGTGGGAGCCGTCCT GTGCAATGTAGGACATTTAGCAGCATCGTTGGCCTCTACCCACTAAAAAC GCCAAGTAGTAAGGCCCCATCCCTTAGTGACAACCCAAAATGTCTCCAGA CATTGCCAATTGCCTCTGGTTGAGACCCACTGATTTATGGAAATCAAAAG AATAATCATTTCCAAAAGCCTGACAGCAAACAGCAAAGTCCAAAAATAAA ATGAGAACAAGACCATTGGCTTTGGTGGTTGGAGGTCATGAGATACCTTC AAGAAAGCACCCTCTATAATATCAAGTCCATGTAGAAGACGTTACAGAGG AAACAAACAATATAAAAGTGAAAACAACCAAGGGTGAGCTACTCTCAGAA AGTATGCCTTTGAAAAGAAAGTCAGAAGCCATAGCTTGGGATCTCTGCAG ATCCCTAAAAGAATAGATTCCTATTCTACTGACTTTCTATGAAGATCAAA TTGTAAAAAGCAAAAGTTTCTCTCCAAGGGTTTCCTTTGCAGTGACCTGT ATGTCCAACCACGCAAGGGCCCATTGTGGGGACATATGTTGTCCAAAAGG ACCATAGCAGAGACAGGCCAGTGAGCCAAAGTGTGGAAACTTTTGAGACT GGCTTGAGCTTGGCACTTATAGAACAATAAACCAAGCCTTTGAAGGGGTT CAACAAAGGAACCATTTGTCCACTCTAGTAGCTACAAAGTAAGGCAGGGT TGCAGCAAAGAACAAAAAAATAAAAGAAGGCCAAGCTGGAGGTATGACCA AAGTTTACTAGGTCCATTCTGAGACCTTCTGCTAGGGTCTGAGATCTAGA AGACAGTGAATAAGGAAACAAACCCAAAACTCAACGCAACACAGGATATG GAAGCTCTCAGGCCTGACGTTAACAGCATCTACTATTTTTCTTCTCAGCT ACTTTAATGAATGCAGTATACTAAAAGCCAGGAGGGGAAGGGACAACACT AAGCAAAAAACATGCATTTTTTAAAATGCACAGATTTTCTTCACTGCCGT TTTTGTTATCATTCCTATGAATTAGTGATGCCGAATTTCATTTTCTCATC TGCTGAAGAGCTTTCCTGTGTTCCTCTCGTTGGAACACATGCTTGGCATT AAAATGCTTGTGAGAACTTCTCTTCCTTTAACGTTCCCTGGCTAGCTTGG TTTTTAATCTAACAGCCCTTCTTTCAAAATGATCCTTCCACTGGAGATAG ATATTTATCATTCTCTTCCTTCACCTCATCTCTTGACAGGCCGCACGTGA CTTGAAGGAATTTTTCAAATAGCAGCTCAGCCACCCTGAGGGGCTTCAGT CTCACCCCTAAGTTCGCTGGCTTTTTCTTCACCACGTCCAGTTGCTTTCC ATCTTATTAACTGCTCTTTTCACTAGAGGACCAACTCAGTAGGAAATTTT TTGAGAGGTGGAGAAAGAGATGTTCAAAGAAGGTGTTGGGGTCGGGGGAA ACTGGTTTTATTTTATACAAGTCACACATTCTGAATCTTCCCTTTTGTGT CTCTGGGGAGAAAGGAGAAAGTTTGATCAAATCGCTCATTATTTCTGCAC TTCTTTCTTTTTTCCTAAGTATAAAAATATATGACTACTACTACTGTGAG ACTATGTGATTGTGAGAATGAATGATTCTTTTTTTTTTTTTTTTTTTTTT GAAACGGAGTCTCTCGCTGTCACCCAGGCTGGAGTGCAGTAGCACGATCT TGGCTCACTGCAACATCTGCCTCCCGGGTTCAAGCAATTCTCCTGCCTCA GCCTCCTGAGTAGCTGGGACTACAGGTGCGCTCCACCACCCCCAGCTAAT TTTTGTATTTTTAGTGGAGACGGGGTTTCACCATGTTGGTCAGGCTGGTC TTGAACTCCTGACCTCATGATCCTCTCACCTCGGCCTCACAAAGTGCTAG GATTACAGGCGCATGGCCAAGAATGAATGATTATTTGTGCCTTCCTATGT GAAAAAAAAATGTTTCCTCTAGCTACACACTATTCTGTTCTGTGAGGCCG CCCCATCAGACTGTTGACCTAGAGTCCCAACCCCGGCCCTCCAGGAGACC TGCCTGTTCTTAGAAGCCCAACCCACTCAGCAGCAGCTCCAAATAACAGG GGGAGCCAACAAAAAAGAGTGCTGCTAGAGCAACAAGCAAGGGGCAATTA GTCAGAAGGCAACTTCCATGGTCTTCCAAAAAAAATTGAGGTGAAAGACC AAAGATGTCCCTAAAATGTCTTCCTAAAAGATAAACTTCATCAACTACCT CTGACTGGTCAGTATTAAGAACCACTTTCAGGCCAGGTGTCATGGTTCAC GCCTGTAACTCCATCTACTCCAGAGGCTGAGGCAGGACAATTGCTTCAGG CCGGAGGATTGCTTGAGGCCAGGAGCTGGAGACCAAGCCTGAGCAACACA GTGAGACCTCATCTCTACCAAAAATGTACCTCTATTAAAAAACAAAAAAG AAGAAGAAGAAGAAGAAGAAGGAGAGGAGGCTGGGTATGGTGGCTAATGC CTTTGTAATCCCAGAACTTTGGAAGGCTGAGGCAGGAGAATCACTTAGGC TGAGGCAGGAGAATCACCAGAGTCTAGGAGTTTGAGACCAGCCTGGGCAA CATAGTGAGACCCCCATCTCTACAAAAAAAAAAATTCAAAAATTAGCCAA GCGTGGGGTTTGTGCCTGTAGACCCAACTACTCAGGAGGCTCAGGTAGGA GGATCACCTGAGTCCAGGGAGGTCGAGGCTGCAGTGAGTCATGATTATTC CACTGCCTTCCAGCCTAGACTACAGGGTGAGACCCTGTCTTAAAAAAAAA ATTAAAGAAGAAAAAACTCTCTTTTCTTTTCTTTCTTTCTTCTTCTTCTT TTCTTTTTTTTTTTTTCTTTTTTTTTTTTTAGAGATGGCACGTCACCACA TTGCCCAGGCTGTTGTCGAACTCCTGGCCTCAAACGATGCTCCCACTTGA GCCTCCCAAAGTGCTGGGACTACAAGCATAAGCCACCACACACGGCCTTT TCCTTTCTTTTTCTATTTCTCAATGGATTTTTCCAATGGACACGTATCAC TTTGGTAGTTATACATGATACTAGTTGTAATCTCAGCCATTTTTCAACCC AGCAAATGTCTATTCTAGGTCAAATATGTCTCAAAAATTACTAAAAGAAA ATCAGTTATGTCCTTTAACCTGGCTGAGGTCTGGCTTTGTTTTCTCTCAT GTAAAAATGGAGATGGCACAAAACAACTCCAAGCTGTTACTTGAAAGTAA CACCTCAGGTGATGTCACCAGCCTGAGGGAGAGTGAGGTTAAGTTCTGAA CCCACAGGCATTATATCTGCCTGGGGTTCACATGCCCTACACTGGACTGG CATAATTTGAGAGTCAGATCCGAAGATGTGGTATATCCGCCATCTTTAGC AACTTTCAAAAACTACCCTATGAGGTCAAGCTGGACCTACTTTTGGTTTT GCCATTGTTGTTTGTTTGTTGTTGAGGGTTTTCTTTGAGGGGCGGGGAGT GCATGCCCCTGTGGAGAGCACTCATTTAGCTTCAATTAGAGTAATGCCAA AAGTGCCAGATTCCTGGGAAATCAGCCTACAAGGCTCCTGCGGGAAGGAA CCTCCACTGCCAGAAGTCCTTAGGGCATCTAAGTGATCAGACACCGTCAG GGATTCTTTGCCCCGTAAAAACCTACTTGACCAGGGACACGTGCCAGGTA AATTTCCTTCACATTTACTTCAACCTTATTGCATACTCATTTTAGTATTA AAACCTTTAATAAAATGCTCCTATTCCTTCACACTTTTTTTCTATGAGAT CTCAAATACCCCTTCTTGCTATTAAAAAAAATCACTTATTATTCACCAGC CCAATATTTTAAAAGTAAAAATAATAAGCCAAGGCCAGGAGCGATGACTC GCACTTGTATTCCCAGCAGTTTCAGAGGCAAAGGCCGAAGGATCGCTTTA ACCGAGGAGTTTGAGACCAGCCTGGGCAACATGACCAGACTGCCTCTCTA CAAAAAGTTTAAAAAATTAACCGGGTGTGGTGGTGCACTGCACTCCCAGC TACTGGGCTGGGGTATCAGGCTGAGGTAGGAGGTTTGCTTTGAGCCCGGG GGGATCGAGGCTGCAGTGAGCTTTGATTGTGCCACTGCACTCCAGCCTGG GTGACAGAAGGAGACCCTGTCTCAAAAATAATAAGAATAATAATTAATAA TAATAGGCCAAACCAAATACCCATCACCTTCTGCTGTGCCTCCCCTTTCC CCAATAAATCCAGTGTCTTGCTTTCAAATTTTGTGGTTAAAAAAGATGAT GAGTTTCTAAGACGTGGGGGCTAAAGCTTGTTTGGCCGTTTTAGGGTTTG TTGGAATTTTTTTTTCGTCTATGTACTTGTGAATTATTTCACGTTTGCCA TTACCGGTTCTCCATAGGGTGATGTTCATTAGCAGTGGTGATAGGTTAAT TTTCACCATCTCTTATGCGGTTGAATAGTCACCTCTGAACCACTTTTTCC TCCAGTAACTCCTCTTTCTTCGGACCTTCTGCAGCCAACCTGAAAGAATA ACAAGGAGGTGGCTGGAAACTTGTTTTAAGGAACCGCCTGTCCTTCCCCC GCTGGAAACCTTGCACCTCGGACGCTCCTGCTCCTGCCCCCACCTGACCC CCGCCCTCGTTGACATCCAGGCGCGATGATCTCTGCTGCCAGTAGAGGGC ACACTTACTTTACTTTCGCAAACCTGAACGCGGGTGCTGCCCAGAGAGGG GGCGGAGGGAAAGACGCTTTGCAGCAAAATCCAGCATAGCGATTGGTTGC TCCCCGCGTTTGCGGCAAAGGCCTGGAGGCAGGAGTAATTTGCAATCCTT AAAGCTGAATTGTGCAGTGCATCGGATTTGGAAGCTACTATATTCACTTA ACACTTGAACGCTGAGCTGCAAACTCAACGGGTAATAACCCATCTTGAAC AGCGTACATGCTATACACGCACCCCTTTCCCCCGAATTGTTTTCTCTTTT GGAGGTGGTGGAGGGAGAGAAAAGTTTACTTAAAATGCCTTTGGGTGAGG GACCAAGGATGAGAAGAATGTTTTTTGTTTTTCATGCCGTGGAATAACAC AAAATAAAAAATCCCGAGGGAATATACATTATATATTAAATATAGATCAT TTCAGGGAGCAAACAAATCATGTGTGGGGCTGGGCAACTAGCTAAGTCGA AGCGTAAATAAAATGTGAATACACGTTTGCGGGTTACATACAGTGCACTT TCACTAGTATTCAGAAAAAATTGTGAGTCAGTGAACTAGGAAATTAATGC CTGGAAGGCAGCCAAATTTTAATTAGCTCAAGACTCCCCCCCCCCCAAAA AAAGGCACGGAAGTAATACTCCTCTCCTCTTCTTTGATCAGAATCGATGC ATTTTTTGTGCATGACCGCATTTCCAATAATAAAAGGGGAAAGAGGACCT GGAAAGGAATTAAACGTCCGGTTTGTCCGGGGAGGAAAGAGTTAACGGTT TTTTTCACAAGGGTCTCTGCTGACTCCCCCGGCTCGGTCCACAAGCTCTC CACTTGCCCCTTTTAGGAAGTCCGGTCCCGCGGTTCGGGTACCCCCTGCC CCTCCCATATTCTCCCGTCTAGCACCTTTGATTTCTCCCAAACCCGGCAG CCCGAGACTGTTGCAAACCGGCGCCACAGGGCGCAAAGGGGATTTGTCTC TTCTGAAACCTGGCTGAGAAATTGGGAACTCCGTGTGGGAGGCGTGGGGG TGGGACGGTGGGGTACAGACTGGCAGAGAGCAGGCAACCTCCCTCTCGCC CTAGCCCAGCTCTGGAACAGGCAGACACATCTCAGGGCTAAACAGACGCC TCCCGCACGGGGCCCCACGGAAGCCTGAGCAGGCGGGGCAGGAGGGGCGG TATCTGCTGCTTTGGCAGCAAATTGGGGGACTCAGTCTGGGTGGAAGGTA TCCAATCCAGATAGCTGTGCATACATAATGCATAATACATGACTCCCCCC AACAAATGCAATGGGAGTTTATTCATAACGCGCTCTCCAAGTATACGTGG CAATGCGTTGCTGGGTTATTTTAATCATTCTAGGCATCGTTTTCCTCCTT ATGCCTCTATCATTCCTCCCTATCTACACTAACATCCCACGCTCTGAACG CGCGCCCATTAATACCCTTCTTTCCTCCACTCTCCCTGGGACTCTTGATC AAAGCGCGGCCCTTTCCCCAGCCTTAGCGAGGCGCCCTGCAGCCTGGTAC GCGCGTGGCGTGGCGGTGGGCGCGCAGTGCGTTCTCGGTGTGGAGGGCAG CTGTTCCGCCTGCGATGATTTATACTCACAGGACAAGGATGCGGTTTGTC AAACAGTACTGCTACGGAGGAGCAGCAGAGAAAGGGAGAGGGTTTGAGAG GGAGCAAAAGAAAATGGTAGGCGCGCGTAGTTAATTCATGCGGCTCTCTT ACTCTGTTTACATCCTAGAGCTAGAGTGCTCGGCTGCCCGGCTGAGTCTC CTCCCCACCTTCCCCACCCTCCCCACCCTCCCCATAAGCGCCCCTCCCGG GTTCCCAAAGCAGAGGGCGTGGGGGAAAAGAAAAAAGATCCTCTCTCGCT AATCTCCGCCCACCGGCCCTTTATAATGCGAGGGTCTGGACGGCTGAGGA CCCCCGAGCTGTGCTGCTCGCGGCCGCCACCGCCGGGCCCCGGCCGTCCC TGGCTCCCCTCCTGCCTCGAGAAGGGCAGGGCTTCTCAGAGGCTTGGCGG GAAAAAGAACGGAGGGAGGGATCGCGCTGAGTATAAAAGCCGGTTTTCGG GGCTTTATCTAACTCGCTGTAGTAATTCCAGCGAGAGGCAGAGGGAGCGA GCGGGCGGCCGGCTAGGGTGGAAGAGCCGGGCGAGCAGAGCTGCGCTGCG GGCGTCCTGGGAAGGGAGATCCGGAGCGAATAGGGGGCTTCGCCTCTGGC CCAGCCCTCCCGCTGATCCCCCAGCCAGCGGTCCGCAACCCTTGCCGCAT CCACGAAACTTTGCCCATAGCAGCGGGCGGGCACTTTGCACTGGAACTTA CAACACCCGAGCAAGGACGCGACTCTCCCGACGCGGGGAGGCTATTCTGC CCATTTGGGGACACTTCCCCGCCGCTGCCAGGACCCGCTTCTCTGAAAGG CTCTCCTTGCAGCTGCTTAGACG CTG

31. APP

Amyloid beta (A4) precursor protein is encoded by the APP gene. The amyloid precursor protein (APP) is found in many tissues and organs, including the brain and spinal cord (central nervous system). Its function is not well understood, however, it is believed to bind other proteins on the surface of cells or help cells attach to one another, thereby directing the migration of nerve cells during early development. APP is cleaved by enzymes to create smaller peptides (soluble amyloid precursor protein (sAPP) and amyloid beta (β) peptide) which may be released outside the cell. sAPP has growth-promoting properties and may play a role in the formation of nerve cells (neurons) in the brain both before and after birth. The sAPP peptide may also control the function of certain other proteins by turning off (inhibiting) their activity. Alzheimer's disease (AD) pathogenesis is widely believed to be driven by the production and deposition of the amyloid-beta peptide (Murphy and Levin (2010) J Alzheimers Dis. 19(1):311-23).

Protein: Beta Amyloid Gene: APP (Homo sapiens, chromosome 21, 27252861-27543446 [NCBI Reference Sequence: NC_(—)000021.8]; start site location: 27542938; strand: negative)

Gene Identification GeneID 351 HGNC 620 MIM 104760

Targeted Sequences Relative upstream Sequence Design location to gene start ID No: ID Sequence (5′-3′) site 7607 CGCGACCCTGCGCGGGGCACCG 1 7741 GTGCGAGTGGGATCCGCCGCG 34 7875 CGCGCCGCCACCGCCGCCGTCTCCCGG 68 8009 CGCGCACGCTCCTCCGCGTGCTCTCG 101 8143 CCGAGGAAACTGACGGAGCCCGAGCGCGG 137 8145 CGAGTCAGCTGATCCGGCCCACCCCG 186 8310 CGAGAGAGACCCCTAGCGGCGCCG 221 8475 CGCCCGCTCGCGCCGGGAGGGGCCCTCG 256 8640 CGCGCCCACAGGTGCACGCGCCCTTGGCG 289 8805 GGCCGACGGCCCACCTGGGCTTCG 351 8825 CGCTGAGGCTCTAGAAAAGTCGAGAG 446 8843 CTCGTCCCCGTGAGCTTGAATCATCCGACCC 480 8912 AGGCGTTTCTGGAAGAGAATGAGAACG 604 8927 CGTCAAAAGCAGGCACGAGCAACCTG 701 8928 GAACGAACCAAAGGAGCAAGGCG 742 8929 CGCTGACAAGGGTGCCTAGGCCCGG 1318 8948 CGCAATTCCGTATTTGTTCCGG 1738 8969 GTACGTTGGCAGACGCAGTGACG 4923

Target Shift Sequences Relative upstream location to Sequence gene start ID No: Sequence (5′-3′) site 7607 CGCGACCCTGCGCGGGGCACCG 1 7608 GCGACCCTGCGCGGGGCACC 2 7609 CGACCCTGCGCGGGGCACCG 3 7610 GACCCTGCGCGGGGCACCGA 4 7611 ACCCTGCGCGGGGCACCGAG 5 7612 CCCTGCGCGGGGCACCGAGT 6 7613 CCTGCGCGGGGCACCGAGTG 7 7614 CTGCGCGGGGCACCGAGTGC 8 7615 TGCGCGGGGCACCGAGTGCG 9 7616 GCGCGGGGCACCGAGTGCGC 10 7617 CGCGGGGCACCGAGTGCGCT 11 7618 GCGGGGCACCGAGTGCGCTG 12 7619 CGGGGCACCGAGTGCGCTGC 13 7620 GGGGCACCGAGTGCGCTGCT 14 7621 GGGCACCGAGTGCGCTGCTG 15 7622 GGCACCGAGTGCGCTGCTGT 16 7623 GCACCGAGTGCGCTGCTGTG 17 7624 CACCGAGTGCGCTGCTGTGC 18 7625 ACCGAGTGCGCTGCTGTGCG 19 7626 CCGAGTGCGCTGCTGTGCGA 20 7627 CGAGTGCGCTGCTGTGCGAG 21 7628 GAGTGCGCTGCTGTGCGAGT 22 7629 AGTGCGCTGCTGTGCGAGTG 23 7630 GTGCGCTGCTGTGCGAGTGG 24 7631 TGCGCTGCTGTGCGAGTGGG 25 7632 GCGCTGCTGTGCGAGTGGGA 26 7633 CGCTGCTGTGCGAGTGGGAT 27 7634 GCTGCTGTGCGAGTGGGATC 28 7635 CTGCTGTGCGAGTGGGATCC 29 7636 TGCTGTGCGAGTGGGATCCG 30 7637 GCTGTGCGAGTGGGATCCGC 31 7638 CTGTGCGAGTGGGATCCGCC 32 7639 TGTGCGAGTGGGATCCGCCG 33 7640 GTGCGAGTGGGATCCGCCGC 34 7641 TGCGAGTGGGATCCGCCGCG 35 7642 GCGAGTGGGATCCGCCGCGT 36 7643 CGAGTGGGATCCGCCGCGTC 37 7644 GAGTGGGATCCGCCGCGTCC 38 7645 AGTGGGATCCGCCGCGTCCT 39 7646 GTGGGATCCGCCGCGTCCTT 40 7647 TGGGATCCGCCGCGTCCTTG 41 7648 GGGATCCGCCGCGTCCTTGC 42 7649 GGATCCGCCGCGTCCTTGCT 43 7650 GATCCGCCGCGTCCTTGCTC 44 7651 ATCCGCCGCGTCCTTGCTCT 45 7652 TCCGCCGCGTCCTTGCTCTG 46 7653 CCGCCGCGTCCTTGCTCTGC 47 7654 CGCCGCGTCCTTGCTCTGCC 48 7655 GCCGCGTCCTTGCTCTGCCC 49 7656 CCGCGTCCTTGCTCTGCCCG 50 7657 CGCGTCCTTGCTCTGCCCGC 51 7658 GCGTCCTTGCTCTGCCCGCG 52 7659 CGTCCTTGCTCTGCCCGCGC 53 7660 GTCCTTGCTCTGCCCGCGCC 54 7661 TCCTTGCTCTGCCCGCGCCG 55 7662 CCTTGCTCTGCCCGCGCCGC 56 7663 CTTGCTCTGCCCGCGCCGCC 57 7664 TTGCTCTGCCCGCGCCGCCA 58 7665 TGCTCTGCCCGCGCCGCCAC 59 7666 GCTCTGCCCGCGCCGCCACC 60 7667 CTCTGCCCGCGCCGCCACCG 61 7668 TCTGCCCGCGCCGCCACCGC 62 7669 CTGCCCGCGCCGCCACCGCC 63 7670 TGCCCGCGCCGCCACCGCCG 64 7671 GCCCGCGCCGCCACCGCCGC 65 7672 CCCGCGCCGCCACCGCCGCC 66 7673 CCGCGCCGCCACCGCCGCCG 67 7674 CGCGCCGCCACCGCCGCCGT 68 7675 GCGCCGCCACCGCCGCCGTC 69 7676 CGCCGCCACCGCCGCCGTCT 70 7677 GCCGCCACCGCCGCCGTCTC 71 7678 CCGCCACCGCCGCCGTCTCC 72 7679 CGCCACCGCCGCCGTCTCCC 73 7680 GCCACCGCCGCCGTCTCCCG 74 7681 CCACCGCCGCCGTCTCCCGG 75 7682 CACCGCCGCCGTCTCCCGGG 76 7683 ACCGCCGCCGTCTCCCGGGG 77 7684 CCGCCGCCGTCTCCCGGGGC 78 7685 CGCCGCCGTCTCCCGGGGCC 79 7686 GCCGCCGTCTCCCGGGGCCC 80 7687 CCGCCGTCTCCCGGGGCCCC 81 7688 CGCCGTCTCCCGGGGCCCCC 82 7689 GCCGTCTCCCGGGGCCCCCG 83 7690 CCGTCTCCCGGGGCCCCCGC 84 7691 CGTCTCCCGGGGCCCCCGCG 85 7692 GTCTCCCGGGGCCCCCGCGC 86 7693 TCTCCCGGGGCCCCCGCGCA 87 7694 CTCCCGGGGCCCCCGCGCAC 88 7695 TCCCGGGGCCCCCGCGCACG 89 7696 CCCGGGGCCCCCGCGCACGC 90 7697 CCGGGGCCCCCGCGCACGCT 91 7698 CGGGGCCCCCGCGCACGCTC 92 7699 GGGGCCCCCGCGCACGCTCC 93 7700 GGGCCCCCGCGCACGCTCCT 94 7701 GGCCCCCGCGCACGCTCCTC 95 7702 GCCCCCGCGCACGCTCCTCC 96 7703 CCCCCGCGCACGCTCCTCCG 97 7704 CCCCGCGCACGCTCCTCCGC 98 7705 CCCGCGCACGCTCCTCCGCG 99 7706 CCGCGCACGCTCCTCCGCGT 100 7707 CGCGCACGCTCCTCCGCGTG 101 7708 GCGCACGCTCCTCCGCGTGC 102 7709 CGCACGCTCCTCCGCGTGCT 103 7710 GCACGCTCCTCCGCGTGCTC 104 7711 CACGCTCCTCCGCGTGCTCT 105 7712 ACGCTCCTCCGCGTGCTCTC 106 7713 CGCTCCTCCGCGTGCTCTCG 107 7714 GCTCCTCCGCGTGCTCTCGC 108 7715 CTCCTCCGCGTGCTCTCGCC 109 7716 TCCTCCGCGTGCTCTCGCCT 110 7717 CCTCCGCGTGCTCTCGCCTA 111 7718 CTCCGCGTGCTCTCGCCTAC 112 7719 TCCGCGTGCTCTCGCCTACC 113 7720 CCGCGTGCTCTCGCCTACCG 114 7721 CGCGTGCTCTCGCCTACCGC 115 7722 GCGTGCTCTCGCCTACCGCT 116 7723 CGTGCTCTCGCCTACCGCTG 117 7724 GTGCTCTCGCCTACCGCTGC 118 7725 TGCTCTCGCCTACCGCTGCC 119 7726 GCTCTCGCCTACCGCTGCCG 120 7727 CTCTCGCCTACCGCTGCCGA 121 7728 TCTCGCCTACCGCTGCCGAG 122 7729 CTCGCCTACCGCTGCCGAGG 123 7730 TCGCCTACCGCTGCCGAGGA 124 7731 CGCCTACCGCTGCCGAGGAA 125 7732 GCCTACCGCTGCCGAGGAAA 126 7733 CCTACCGCTGCCGAGGAAAC 127 7734 CTACCGCTGCCGAGGAAACT 128 7735 TACCGCTGCCGAGGAAACTG 129 7736 ACCGCTGCCGAGGAAACTGA 130 7737 CCGCTGCCGAGGAAACTGAC 131 7738 CGCTGCCGAGGAAACTGACG 132 7739 GCTGCCGAGGAAACTGACGG 133 7740 CTGCCGAGGAAACTGACGGA 134 7741 GTGCGAGTGGGATCCGCCGCG 34 7742 TGCGAGTGGGATCCGCCGCG 35 7743 GCGAGTGGGATCCGCCGCGT 36 7744 CGAGTGGGATCCGCCGCGTC 37 7745 GAGTGGGATCCGCCGCGTCC 38 7746 AGTGGGATCCGCCGCGTCCT 39 7747 GTGGGATCCGCCGCGTCCTT 40 7748 TGGGATCCGCCGCGTCCTTG 41 7749 GGGATCCGCCGCGTCCTTGC 42 7750 GGATCCGCCGCGTCCTTGCT 43 7751 GATCCGCCGCGTCCTTGCTC 44 7752 ATCCGCCGCGTCCTTGCTCT 45 7753 TCCGCCGCGTCCTTGCTCTG 46 7754 CCGCCGCGTCCTTGCTCTGC 47 7755 CGCCGCGTCCTTGCTCTGCC 48 7756 GCCGCGTCCTTGCTCTGCCC 49 7757 CCGCGTCCTTGCTCTGCCCG 50 7758 CGCGTCCTTGCTCTGCCCGC 51 7759 GCGTCCTTGCTCTGCCCGCG 52 7760 CGTCCTTGCTCTGCCCGCGC 53 7761 GTCCTTGCTCTGCCCGCGCC 54 7762 TCCTTGCTCTGCCCGCGCCG 55 7763 CCTTGCTCTGCCCGCGCCGC 56 7764 CTTGCTCTGCCCGCGCCGCC 57 7765 TTGCTCTGCCCGCGCCGCCA 58 7766 TGCTCTGCCCGCGCCGCCAC 59 7767 GCTCTGCCCGCGCCGCCACC 60 7768 CTCTGCCCGCGCCGCCACCG 61 7769 TCTGCCCGCGCCGCCACCGC 62 7770 CTGCCCGCGCCGCCACCGCC 63 7771 TGCCCGCGCCGCCACCGCCG 64 7772 GCCCGCGCCGCCACCGCCGC 65 7773 CCCGCGCCGCCACCGCCGCC 66 7774 CCGCGCCGCCACCGCCGCCG 67 7775 CGCGCCGCCACCGCCGCCGT 68 7776 GCGCCGCCACCGCCGCCGTC 69 7777 CGCCGCCACCGCCGCCGTCT 70 7778 GCCGCCACCGCCGCCGTCTC 71 7779 CCGCCACCGCCGCCGTCTCC 72 7780 CGCCACCGCCGCCGTCTCCC 73 7781 GCCACCGCCGCCGTCTCCCG 74 7782 CCACCGCCGCCGTCTCCCGG 75 7783 CACCGCCGCCGTCTCCCGGG 76 7784 ACCGCCGCCGTCTCCCGGGG 77 7785 CCGCCGCCGTCTCCCGGGGC 78 7786 CGCCGCCGTCTCCCGGGGCC 79 7787 GCCGCCGTCTCCCGGGGCCC 80 7788 CCGCCGTCTCCCGGGGCCCC 81 7789 CGCCGTCTCCCGGGGCCCCC 82 7790 GCCGTCTCCCGGGGCCCCCG 83 7791 CCGTCTCCCGGGGCCCCCGC 84 7792 CGTCTCCCGGGGCCCCCGCG 85 7793 GTCTCCCGGGGCCCCCGCGC 86 7794 TCTCCCGGGGCCCCCGCGCA 87 7795 CTCCCGGGGCCCCCGCGCAC 88 7796 TCCCGGGGCCCCCGCGCACG 89 7797 CCCGGGGCCCCCGCGCACGC 90 7798 CCGGGGCCCCCGCGCACGCT 91 7799 CGGGGCCCCCGCGCACGCTC 92 7800 GGGGCCCCCGCGCACGCTCC 93 7801 GGGCCCCCGCGCACGCTCCT 94 7802 GGCCCCCGCGCACGCTCCTC 95 7803 GCCCCCGCGCACGCTCCTCC 96 7804 CCCCCGCGCACGCTCCTCCG 97 7805 CCCCGCGCACGCTCCTCCGC 98 7806 CCCGCGCACGCTCCTCCGCG 99 7807 CCGCGCACGCTCCTCCGCGT 100 7808 CGCGCACGCTCCTCCGCGTG 101 7809 GCGCACGCTCCTCCGCGTGC 102 7810 CGCACGCTCCTCCGCGTGCT 103 7811 GCACGCTCCTCCGCGTGCTC 104 7812 CACGCTCCTCCGCGTGCTCT 105 7813 ACGCTCCTCCGCGTGCTCTC 106 7814 CGCTCCTCCGCGTGCTCTCG 107 7815 GCTCCTCCGCGTGCTCTCGC 108 7816 CTCCTCCGCGTGCTCTCGCC 109 7817 TCCTCCGCGTGCTCTCGCCT 110 7818 CCTCCGCGTGCTCTCGCCTA 111 7819 CTCCGCGTGCTCTCGCCTAC 112 7820 TCCGCGTGCTCTCGCCTACC 113 7821 CCGCGTGCTCTCGCCTACCG 114 7822 CGCGTGCTCTCGCCTACCGC 115 7823 GCGTGCTCTCGCCTACCGCT 116 7824 CGTGCTCTCGCCTACCGCTG 117 7825 GTGCTCTCGCCTACCGCTGC 118 7826 TGCTCTCGCCTACCGCTGCC 119 7827 GCTCTCGCCTACCGCTGCCG 120 7828 CTCTCGCCTACCGCTGCCGA 121 7829 TCTCGCCTACCGCTGCCGAG 122 7830 CTCGCCTACCGCTGCCGAGG 123 7831 TCGCCTACCGCTGCCGAGGA 124 7832 CGCCTACCGCTGCCGAGGAA 125 7833 GCCTACCGCTGCCGAGGAAA 126 7834 CCTACCGCTGCCGAGGAAAC 127 7835 CTACCGCTGCCGAGGAAACT 128 7836 TACCGCTGCCGAGGAAACTG 129 7837 ACCGCTGCCGAGGAAACTGA 130 7838 CCGCTGCCGAGGAAACTGAC 131 7839 CGCTGCCGAGGAAACTGACG 132 7840 GCTGCCGAGGAAACTGACGG 133 7841 CTGCCGAGGAAACTGACGGA 134 7842 TGTGCGAGTGGGATCCGCCG 33 7843 CTGTGCGAGTGGGATCCGCC 32 7844 GCTGTGCGAGTGGGATCCGC 31 7845 TGCTGTGCGAGTGGGATCCG 30 7846 CTGCTGTGCGAGTGGGATCC 29 7847 GCTGCTGTGCGAGTGGGATC 28 7848 CGCTGCTGTGCGAGTGGGAT 27 7849 GCGCTGCTGTGCGAGTGGGA 26 7850 TGCGCTGCTGTGCGAGTGGG 25 7851 GTGCGCTGCTGTGCGAGTGG 24 7852 AGTGCGCTGCTGTGCGAGTG 23 7853 GAGTGCGCTGCTGTGCGAGT 22 7854 CGAGTGCGCTGCTGTGCGAG 21 7855 CCGAGTGCGCTGCTGTGCGA 20 7856 ACCGAGTGCGCTGCTGTGCG 19 7857 CACCGAGTGCGCTGCTGTGC 18 7858 GCACCGAGTGCGCTGCTGTG 17 7859 GGCACCGAGTGCGCTGCTGT 16 7860 GGGCACCGAGTGCGCTGCTG 15 7861 GGGGCACCGAGTGCGCTGCT 14 7862 CGGGGCACCGAGTGCGCTGC 13 7863 GCGGGGCACCGAGTGCGCTG 12 7864 CGCGGGGCACCGAGTGCGCT 11 7865 GCGCGGGGCACCGAGTGCGC 10 7866 TGCGCGGGGCACCGAGTGCG 9 7867 CTGCGCGGGGCACCGAGTGC 8 7868 CCTGCGCGGGGCACCGAGTG 7 7869 CCCTGCGCGGGGCACCGAGT 6 7870 ACCCTGCGCGGGGCACCGAG 5 7871 GACCCTGCGCGGGGCACCGA 4 7872 CGACCCTGCGCGGGGCACCG 3 7873 GCGACCCTGCGCGGGGCACC 2 7874 CGCGACCCTGCGCGGGGCAC 1 7875 CGCGCCGCCACCGCCGCCGTCTCCCGG 68 7876 GCGCCGCCACCGCCGCCGTC 69 7877 CGCCGCCACCGCCGCCGTCT 70 7878 GCCGCCACCGCCGCCGTCTC 71 7879 CCGCCACCGCCGCCGTCTCC 72 7880 CGCCACCGCCGCCGTCTCCC 73 7881 GCCACCGCCGCCGTCTCCCG 74 7882 CCACCGCCGCCGTCTCCCGG 75 7883 CACCGCCGCCGTCTCCCGGG 76 7884 ACCGCCGCCGTCTCCCGGGG 77 7885 CCGCCGCCGTCTCCCGGGGC 78 7886 CGCCGCCGTCTCCCGGGGCC 79 7887 GCCGCCGTCTCCCGGGGCCC 80 7888 CCGCCGTCTCCCGGGGCCCC 81 7889 CGCCGTCTCCCGGGGCCCCC 82 7890 GCCGTCTCCCGGGGCCCCCG 83 7891 CCGTCTCCCGGGGCCCCCGC 84 7892 CGTCTCCCGGGGCCCCCGCG 85 7893 GTCTCCCGGGGCCCCCGCGC 86 7894 TCTCCCGGGGCCCCCGCGCA 87 7895 CTCCCGGGGCCCCCGCGCAC 88 7896 TCCCGGGGCCCCCGCGCACG 89 7897 CCCGGGGCCCCCGCGCACGC 90 7898 CCGGGGCCCCCGCGCACGCT 91 7899 CGGGGCCCCCGCGCACGCTC 92 7900 GGGGCCCCCGCGCACGCTCC 93 7901 GGGCCCCCGCGCACGCTCCT 94 7902 GGCCCCCGCGCACGCTCCTC 95 7903 GCCCCCGCGCACGCTCCTCC 96 7904 CCCCCGCGCACGCTCCTCCG 97 7905 CCCCGCGCACGCTCCTCCGC 98 7906 CCCGCGCACGCTCCTCCGCG 99 7907 CCGCGCACGCTCCTCCGCGT 100 7908 CGCGCACGCTCCTCCGCGTG 101 7909 GCGCACGCTCCTCCGCGTGC 102 7910 CGCACGCTCCTCCGCGTGCT 103 7911 GCACGCTCCTCCGCGTGCTC 104 7912 CACGCTCCTCCGCGTGCTCT 105 7913 ACGCTCCTCCGCGTGCTCTC 106 7914 CGCTCCTCCGCGTGCTCTCG 107 7915 GCTCCTCCGCGTGCTCTCGC 108 7916 CTCCTCCGCGTGCTCTCGCC 109 7917 TCCTCCGCGTGCTCTCGCCT 110 7918 CCTCCGCGTGCTCTCGCCTA 111 7919 CTCCGCGTGCTCTCGCCTAC 112 7920 TCCGCGTGCTCTCGCCTACC 113 7921 CCGCGTGCTCTCGCCTACCG 114 7922 CGCGTGCTCTCGCCTACCGC 115 7923 GCGTGCTCTCGCCTACCGCT 116 7924 CGTGCTCTCGCCTACCGCTG 117 7925 GTGCTCTCGCCTACCGCTGC 118 7926 TGCTCTCGCCTACCGCTGCC 119 7927 GCTCTCGCCTACCGCTGCCG 120 7928 CTCTCGCCTACCGCTGCCGA 121 7929 TCTCGCCTACCGCTGCCGAG 122 7930 CTCGCCTACCGCTGCCGAGG 123 7931 TCGCCTACCGCTGCCGAGGA 124 7932 CGCCTACCGCTGCCGAGGAA 125 7933 GCCTACCGCTGCCGAGGAAA 126 7934 CCTACCGCTGCCGAGGAAAC 127 7935 CTACCGCTGCCGAGGAAACT 128 7936 TACCGCTGCCGAGGAAACTG 129 7937 ACCGCTGCCGAGGAAACTGA 130 7938 CCGCTGCCGAGGAAACTGAC 131 7939 CGCTGCCGAGGAAACTGACG 132 7940 GCTGCCGAGGAAACTGACGG 133 7941 CTGCCGAGGAAACTGACGGA 134 7942 CCGCGCCGCCACCGCCGCCG 67 7943 CCCGCGCCGCCACCGCCGCC 66 7944 GCCCGCGCCGCCACCGCCGC 65 7945 TGCCCGCGCCGCCACCGCCG 64 7946 CTGCCCGCGCCGCCACCGCC 63 7947 TCTGCCCGCGCCGCCACCGC 62 7948 CTCTGCCCGCGCCGCCACCG 61 7949 GCTCTGCCCGCGCCGCCACC 60 7950 TGCTCTGCCCGCGCCGCCAC 59 7951 TTGCTCTGCCCGCGCCGCCA 58 7952 CTTGCTCTGCCCGCGCCGCC 57 7953 CCTTGCTCTGCCCGCGCCGC 56 7954 TCCTTGCTCTGCCCGCGCCG 55 7955 GTCCTTGCTCTGCCCGCGCC 54 7956 CGTCCTTGCTCTGCCCGCGC 53 7957 GCGTCCTTGCTCTGCCCGCG 52 7958 CGCGTCCTTGCTCTGCCCGC 51 7959 CCGCGTCCTTGCTCTGCCCG 50 7960 GCCGCGTCCTTGCTCTGCCC 49 7961 CGCCGCGTCCTTGCTCTGCC 48 7962 CCGCCGCGTCCTTGCTCTGC 47 7963 TCCGCCGCGTCCTTGCTCTG 46 7964 ATCCGCCGCGTCCTTGCTCT 45 7965 GATCCGCCGCGTCCTTGCTC 44 7966 GGATCCGCCGCGTCCTTGCT 43 7967 GGGATCCGCCGCGTCCTTGC 42 7968 TGGGATCCGCCGCGTCCTTG 41 7969 GTGGGATCCGCCGCGTCCTT 40 7970 AGTGGGATCCGCCGCGTCCT 39 7971 GAGTGGGATCCGCCGCGTCC 38 7972 CGAGTGGGATCCGCCGCGTC 37 7973 GCGAGTGGGATCCGCCGCGT 36 7974 TGCGAGTGGGATCCGCCGCG 35 7975 GTGCGAGTGGGATCCGCCGC 34 7976 TGTGCGAGTGGGATCCGCCG 33 7977 CTGTGCGAGTGGGATCCGCC 32 7978 GCTGTGCGAGTGGGATCCGC 31 7979 TGCTGTGCGAGTGGGATCCG 30 7980 CTGCTGTGCGAGTGGGATCC 29 7981 GCTGCTGTGCGAGTGGGATC 28 7982 CGCTGCTGTGCGAGTGGGAT 27 7983 GCGCTGCTGTGCGAGTGGGA 26 7984 TGCGCTGCTGTGCGAGTGGG 25 7985 GTGCGCTGCTGTGCGAGTGG 24 7986 AGTGCGCTGCTGTGCGAGTG 23 7987 GAGTGCGCTGCTGTGCGAGT 22 7988 CGAGTGCGCTGCTGTGCGAG 21 7989 CCGAGTGCGCTGCTGTGCGA 20 7990 ACCGAGTGCGCTGCTGTGCG 19 7991 CACCGAGTGCGCTGCTGTGC 18 7992 GCACCGAGTGCGCTGCTGTG 17 7993 GGCACCGAGTGCGCTGCTGT 16 7994 GGGCACCGAGTGCGCTGCTG 15 7995 GGGGCACCGAGTGCGCTGCT 14 7996 CGGGGCACCGAGTGCGCTGC 13 7997 GCGGGGCACCGAGTGCGCTG 12 7998 CGCGGGGCACCGAGTGCGCT 11 7999 GCGCGGGGCACCGAGTGCGC 10 8000 TGCGCGGGGCACCGAGTGCG 9 8001 CTGCGCGGGGCACCGAGTGC 8 8002 CCTGCGCGGGGCACCGAGTG 7 8003 CCCTGCGCGGGGCACCGAGT 6 8004 ACCCTGCGCGGGGCACCGAG 5 8005 GACCCTGCGCGGGGCACCGA 4 8006 CGACCCTGCGCGGGGCACCG 3 8007 GCGACCCTGCGCGGGGCACC 2 8008 CGCGACCCTGCGCGGGGCAC 1 8009 CGCGCACGCTCCTCCGCGTGCTCTCG 101 8010 GCGCACGCTCCTCCGCGTGC 102 8011 CGCACGCTCCTCCGCGTGCT 103 8012 GCACGCTCCTCCGCGTGCTC 104 8013 CACGCTCCTCCGCGTGCTCT 105 8014 ACGCTCCTCCGCGTGCTCTC 106 8015 CGCTCCTCCGCGTGCTCTCG 107 8016 GCTCCTCCGCGTGCTCTCGC 108 8017 CTCCTCCGCGTGCTCTCGCC 109 8018 TCCTCCGCGTGCTCTCGCCT 110 8019 CCTCCGCGTGCTCTCGCCTA 111 8020 CTCCGCGTGCTCTCGCCTAC 112 8021 TCCGCGTGCTCTCGCCTACC 113 8022 CCGCGTGCTCTCGCCTACCG 114 8023 CGCGTGCTCTCGCCTACCGC 115 8024 GCGTGCTCTCGCCTACCGCT 116 8025 CGTGCTCTCGCCTACCGCTG 117 8026 GTGCTCTCGCCTACCGCTGC 118 8027 TGCTCTCGCCTACCGCTGCC 119 8028 GCTCTCGCCTACCGCTGCCG 120 8029 CTCTCGCCTACCGCTGCCGA 121 8030 TCTCGCCTACCGCTGCCGAG 122 8031 CTCGCCTACCGCTGCCGAGG 123 8032 TCGCCTACCGCTGCCGAGGA 124 8033 CGCCTACCGCTGCCGAGGAA 125 8034 GCCTACCGCTGCCGAGGAAA 126 8035 CCTACCGCTGCCGAGGAAAC 127 8036 CTACCGCTGCCGAGGAAACT 128 8037 TACCGCTGCCGAGGAAACTG 129 8038 ACCGCTGCCGAGGAAACTGA 130 8039 CCGCTGCCGAGGAAACTGAC 131 8040 CGCTGCCGAGGAAACTGACG 132 8041 GCTGCCGAGGAAACTGACGG 133 8042 CTGCCGAGGAAACTGACGGA 134 8043 CCGCGCACGCTCCTCCGCGT 100 8044 CCCGCGCACGCTCCTCCGCG 99 8045 CCCCGCGCACGCTCCTCCGC 98 8046 CCCCCGCGCACGCTCCTCCG 97 8047 GCCCCCGCGCACGCTCCTCC 96 8048 GGCCCCCGCGCACGCTCCTC 95 8049 GGGCCCCCGCGCACGCTCCT 94 8050 GGGGCCCCCGCGCACGCTCC 93 8051 CGGGGCCCCCGCGCACGCTC 92 8052 CCGGGGCCCCCGCGCACGCT 91 8053 CCCGGGGCCCCCGCGCACGC 90 8054 TCCCGGGGCCCCCGCGCACG 89 8055 CTCCCGGGGCCCCCGCGCAC 88 8056 TCTCCCGGGGCCCCCGCGCA 87 8057 GTCTCCCGGGGCCCCCGCGC 86 8058 CGTCTCCCGGGGCCCCCGCG 85 8059 CCGTCTCCCGGGGCCCCCGC 84 8060 GCCGTCTCCCGGGGCCCCCG 83 8061 CGCCGTCTCCCGGGGCCCCC 82 8062 CCGCCGTCTCCCGGGGCCCC 81 8063 GCCGCCGTCTCCCGGGGCCC 80 8064 CGCCGCCGTCTCCCGGGGCC 79 8065 CCGCCGCCGTCTCCCGGGGC 78 8066 ACCGCCGCCGTCTCCCGGGG 77 8067 CACCGCCGCCGTCTCCCGGG 76 8068 CCACCGCCGCCGTCTCCCGG 75 8069 GCCACCGCCGCCGTCTCCCG 74 8070 CGCCACCGCCGCCGTCTCCC 73 8071 CCGCCACCGCCGCCGTCTCC 72 8072 GCCGCCACCGCCGCCGTCTC 71 8073 CGCCGCCACCGCCGCCGTCT 70 8074 GCGCCGCCACCGCCGCCGTC 69 8075 CGCGCCGCCACCGCCGCCGT 68 8076 CCGCGCCGCCACCGCCGCCG 67 8077 CCCGCGCCGCCACCGCCGCC 66 8078 GCCCGCGCCGCCACCGCCGC 65 8079 TGCCCGCGCCGCCACCGCCG 64 8080 CTGCCCGCGCCGCCACCGCC 63 8081 TCTGCCCGCGCCGCCACCGC 62 8082 CTCTGCCCGCGCCGCCACCG 61 8083 GCTCTGCCCGCGCCGCCACC 60 8084 TGCTCTGCCCGCGCCGCCAC 59 8085 TTGCTCTGCCCGCGCCGCCA 58 8086 CTTGCTCTGCCCGCGCCGCC 57 8087 CCTTGCTCTGCCCGCGCCGC 56 8088 TCCTTGCTCTGCCCGCGCCG 55 8089 GTCCTTGCTCTGCCCGCGCC 54 8090 CGTCCTTGCTCTGCCCGCGC 53 8091 GCGTCCTTGCTCTGCCCGCG 52 8092 CGCGTCCTTGCTCTGCCCGC 51 8093 CCGCGTCCTTGCTCTGCCCG 50 8094 GCCGCGTCCTTGCTCTGCCC 49 8095 CGCCGCGTCCTTGCTCTGCC 48 8096 CCGCCGCGTCCTTGCTCTGC 47 8097 TCCGCCGCGTCCTTGCTCTG 46 8098 ATCCGCCGCGTCCTTGCTCT 45 8099 GATCCGCCGCGTCCTTGCTC 44 8100 GGATCCGCCGCGTCCTTGCT 43 8101 GGGATCCGCCGCGTCCTTGC 42 8102 TGGGATCCGCCGCGTCCTTG 41 8103 GTGGGATCCGCCGCGTCCTT 40 8104 AGTGGGATCCGCCGCGTCCT 39 8105 GAGTGGGATCCGCCGCGTCC 38 8106 CGAGTGGGATCCGCCGCGTC 37 8107 GCGAGTGGGATCCGCCGCGT 36 8108 TGCGAGTGGGATCCGCCGCG 35 8109 GTGCGAGTGGGATCCGCCGC 34 8110 TGTGCGAGTGGGATCCGCCG 33 8111 CTGTGCGAGTGGGATCCGCC 32 8112 GCTGTGCGAGTGGGATCCGC 31 8113 TGCTGTGCGAGTGGGATCCG 30 8114 CTGCTGTGCGAGTGGGATCC 29 8115 GCTGCTGTGCGAGTGGGATC 28 8116 CGCTGCTGTGCGAGTGGGAT 27 8117 GCGCTGCTGTGCGAGTGGGA 26 8118 TGCGCTGCTGTGCGAGTGGG 25 8119 GTGCGCTGCTGTGCGAGTGG 24 8120 AGTGCGCTGCTGTGCGAGTG 23 8121 GAGTGCGCTGCTGTGCGAGT 22 8122 CGAGTGCGCTGCTGTGCGAG 21 8123 CCGAGTGCGCTGCTGTGCGA 20 8124 ACCGAGTGCGCTGCTGTGCG 19 8125 CACCGAGTGCGCTGCTGTGC 18 8126 GCACCGAGTGCGCTGCTGTG 17 8127 GGCACCGAGTGCGCTGCTGT 16 8128 GGGCACCGAGTGCGCTGCTG 15 8129 GGGGCACCGAGTGCGCTGCT 14 8130 CGGGGCACCGAGTGCGCTGC 13 8131 GCGGGGCACCGAGTGCGCTG 12 8132 CGCGGGGCACCGAGTGCGCT 11 8133 GCGCGGGGCACCGAGTGCGC 10 8134 TGCGCGGGGCACCGAGTGCG 9 8135 CTGCGCGGGGCACCGAGTGC 8 8136 CCTGCGCGGGGCACCGAGTG 7 8137 CCCTGCGCGGGGCACCGAGT 6 8138 ACCCTGCGCGGGGCACCGAG 5 8139 GACCCTGCGCGGGGCACCGA 4 8140 CGACCCTGCGCGGGGCACCG 3 8141 GCGACCCTGCGCGGGGCACC 2 8142 CGCGACCCTGCGCGGGGCAC 1 8143 CCGAGGAAACTGACGGAGCCCGAGCGCGG 137 8144 CGAGGAAACTGACGGAGCCC 138 8145 CGAGTCAGCTGATCCGGCCCACCCCG 186 8146 GAGTCAGCTGATCCGGCCCA 187 8147 AGTCAGCTGATCCGGCCCAC 188 8148 GTCAGCTGATCCGGCCCACC 189 8149 TCAGCTGATCCGGCCCACCC 190 8150 CAGCTGATCCGGCCCACCCC 191 8151 AGCTGATCCGGCCCACCCCG 192 8152 GCTGATCCGGCCCACCCCGC 193 8153 CTGATCCGGCCCACCCCGCT 194 8154 TGATCCGGCCCACCCCGCTC 195 8155 GATCCGGCCCACCCCGCTCG 196 8156 ATCCGGCCCACCCCGCTCGG 197 8157 TCCGGCCCACCCCGCTCGGC 198 8158 CCGGCCCACCCCGCTCGGCA 199 8159 CGGCCCACCCCGCTCGGCAC 200 8160 GGCCCACCCCGCTCGGCACC 201 8161 GCCCACCCCGCTCGGCACCC 202 8162 CCCACCCCGCTCGGCACCCG 203 8163 CCACCCCGCTCGGCACCCGA 204 8164 CACCCCGCTCGGCACCCGAG 205 8165 ACCCCGCTCGGCACCCGAGA 206 8166 CCCCGCTCGGCACCCGAGAG 207 8167 CCCGCTCGGCACCCGAGAGA 208 8168 CCGCTCGGCACCCGAGAGAG 209 8169 CGCTCGGCACCCGAGAGAGA 210 8170 GCTCGGCACCCGAGAGAGAC 211 8171 CTCGGCACCCGAGAGAGACC 212 8172 TCGGCACCCGAGAGAGACCC 213 8173 CGGCACCCGAGAGAGACCCC 214 8174 GGCACCCGAGAGAGACCCCT 215 8175 GCACCCGAGAGAGACCCCTA 216 8176 CACCCGAGAGAGACCCCTAG 217 8177 ACCCGAGAGAGACCCCTAGC 218 8178 CCCGAGAGAGACCCCTAGCG 219 8179 CCGAGAGAGACCCCTAGCGG 220 8180 CGAGAGAGACCCCTAGCGGC 221 8181 GAGAGAGACCCCTAGCGGCG 222 8182 AGAGAGACCCCTAGCGGCGC 223 8183 GAGAGACCCCTAGCGGCGCC 224 8184 AGAGACCCCTAGCGGCGCCG 225 8185 GAGACCCCTAGCGGCGCCGC 226 8186 AGACCCCTAGCGGCGCCGCC 227 8187 GACCCCTAGCGGCGCCGCCG 228 8188 ACCCCTAGCGGCGCCGCCGG 229 8189 CCCCTAGCGGCGCCGCCGGG 230 8190 CCCTAGCGGCGCCGCCGGGG 231 8191 CCTAGCGGCGCCGCCGGGGA 232 8192 CTAGCGGCGCCGCCGGGGAA 233 8193 TAGCGGCGCCGCCGGGGAAC 234 8194 AGCGGCGCCGCCGGGGAACT 235 8195 GCGGCGCCGCCGGGGAACTG 236 8196 CGGCGCCGCCGGGGAACTGC 237 8197 GGCGCCGCCGGGGAACTGCG 238 8198 GCGCCGCCGGGGAACTGCGC 239 8199 CGCCGCCGGGGAACTGCGCC 240 8200 GCCGCCGGGGAACTGCGCCC 241 8201 CCGCCGGGGAACTGCGCCCG 242 8202 CGCCGGGGAACTGCGCCCGC 243 8203 GCCGGGGAACTGCGCCCGCT 244 8204 CCGGGGAACTGCGCCCGCTC 245 8205 CGGGGAACTGCGCCCGCTCG 246 8206 GGGGAACTGCGCCCGCTCGC 247 8207 GGGAACTGCGCCCGCTCGCG 248 8208 GGAACTGCGCCCGCTCGCGC 249 8209 GAACTGCGCCCGCTCGCGCC 250 8210 AACTGCGCCCGCTCGCGCCG 251 8211 ACTGCGCCCGCTCGCGCCGG 252 8212 CTGCGCCCGCTCGCGCCGGG 253 8213 TGCGCCCGCTCGCGCCGGGA 254 8214 GCGCCCGCTCGCGCCGGGAG 255 8215 CGCCCGCTCGCGCCGGGAGG 256 8216 GCCCGCTCGCGCCGGGAGGG 257 8217 CCCGCTCGCGCCGGGAGGGG 258 8218 CCGCTCGCGCCGGGAGGGGC 259 8219 CGCTCGCGCCGGGAGGGGCC 260 8220 GCTCGCGCCGGGAGGGGCCC 261 8221 CTCGCGCCGGGAGGGGCCCT 262 8222 TCGCGCCGGGAGGGGCCCTC 263 8223 CGCGCCGGGAGGGGCCCTCG 264 8224 GCGCCGGGAGGGGCCCTCGC 265 8225 CGCCGGGAGGGGCCCTCGCG 266 8226 GCCGGGAGGGGCCCTCGCGC 267 8227 CCGGGAGGGGCCCTCGCGCC 268 8228 CGGGAGGGGCCCTCGCGCCC 269 8229 GGGAGGGGCCCTCGCGCCCC 270 8230 GGAGGGGCCCTCGCGCCCCG 271 8231 GAGGGGCCCTCGCGCCCCGC 272 8232 AGGGGCCCTCGCGCCCCGCG 273 8233 GGGGCCCTCGCGCCCCGCGC 274 8234 GGGCCCTCGCGCCCCGCGCC 275 8235 GGCCCTCGCGCCCCGCGCCC 276 8236 GCCCTCGCGCCCCGCGCCCA 277 8237 CCCTCGCGCCCCGCGCCCAC 278 8238 CCTCGCGCCCCGCGCCCACA 279 8239 CTCGCGCCCCGCGCCCACAG 280 8240 TCGCGCCCCGCGCCCACAGG 281 8241 CGCGCCCCGCGCCCACAGGT 282 8242 GCGCCCCGCGCCCACAGGTG 283 8243 CGCCCCGCGCCCACAGGTGC 284 8244 GCCCCGCGCCCACAGGTGCA 285 8245 CCCCGCGCCCACAGGTGCAC 286 8246 CCCGCGCCCACAGGTGCACG 287 8247 CCGCGCCCACAGGTGCACGC 288 8248 CGCGCCCACAGGTGCACGCG 289 8249 GCGCCCACAGGTGCACGCGC 290 8250 CGCCCACAGGTGCACGCGCC 291 8251 GCCCACAGGTGCACGCGCCC 292 8252 CCCACAGGTGCACGCGCCCT 293 8253 CCACAGGTGCACGCGCCCTT 294 8254 CACAGGTGCACGCGCCCTTG 295 8255 ACAGGTGCACGCGCCCTTGG 296 8256 CAGGTGCACGCGCCCTTGGC 297 8257 AGGTGCACGCGCCCTTGGCG 298 8258 GGTGCACGCGCCCTTGGCGC 299 8259 GTGCACGCGCCCTTGGCGCC 300 8260 TGCACGCGCCCTTGGCGCCG 301 8261 GCACGCGCCCTTGGCGCCGC 302 8262 CACGCGCCCTTGGCGCCGCC 303 8263 ACGCGCCCTTGGCGCCGCCT 304 8264 CGCGCCCTTGGCGCCGCCTG 305 8265 GCGCCCTTGGCGCCGCCTGC 306 8266 CGCCCTTGGCGCCGCCTGCA 307 8267 GCCCTTGGCGCCGCCTGCAC 308 8268 CCCTTGGCGCCGCCTGCACC 309 8269 CCTTGGCGCCGCCTGCACCC 310 8270 CTTGGCGCCGCCTGCACCCC 311 8271 TTGGCGCCGCCTGCACCCCA 312 8272 TGGCGCCGCCTGCACCCCAC 313 8273 GGCGCCGCCTGCACCCCACG 314 8274 GCGCCGCCTGCACCCCACGC 315 8275 CGCCGCCTGCACCCCACGCG 316 8276 GCCGCCTGCACCCCACGCGC 317 8277 CCGCCTGCACCCCACGCGCC 318 8278 CGCCTGCACCCCACGCGCCC 319 8279 GCCTGCACCCCACGCGCCCC 320 8280 CCTGCACCCCACGCGCCCCC 321 8281 CTGCACCCCACGCGCCCCCT 322 8282 TGCACCCCACGCGCCCCCTC 323 8283 GCACCCCACGCGCCCCCTCC 324 8284 CACCCCACGCGCCCCCTCCG 325 8285 ACCCCACGCGCCCCCTCCGC 326 8286 CCCCACGCGCCCCCTCCGCT 327 8287 CCCACGCGCCCCCTCCGCTC 328 8288 CCACGCGCCCCCTCCGCTCC 329 8289 CACGCGCCCCCTCCGCTCCC 330 8290 ACGCGCCCCCTCCGCTCCCC 331 8291 CGCGCCCCCTCCGCTCCCCG 332 8292 GCGCCCCCTCCGCTCCCCGG 333 8293 CGCCCCCTCCGCTCCCCGGC 334 8294 GCCCCCTCCGCTCCCCGGCC 335 8295 GCGAGTCAGCTGATCCGGCC 185 8296 GGCGAGTCAGCTGATCCGGC 184 8297 AGGCGAGTCAGCTGATCCGG 183 8298 CAGGCGAGTCAGCTGATCCG 182 8299 CCAGGCGAGTCAGCTGATCC 181 8300 GCCAGGCGAGTCAGCTGATC 180 8301 AGCCAGGCGAGTCAGCTGAT 179 8302 GAGCCAGGCGAGTCAGCTGA 178 8303 AGAGCCAGGCGAGTCAGCTG 177 8304 CAGAGCCAGGCGAGTCAGCT 176 8305 TCAGAGCCAGGCGAGTCAGC 175 8306 CTCAGAGCCAGGCGAGTCAG 174 8307 GCTCAGAGCCAGGCGAGTCA 173 8308 GGCTCAGAGCCAGGCGAGTC 172 8309 GGGCTCAGAGCCAGGCGAGT 171 8310 CGAGAGAGACCCCTAGCGGCGCCG 221 8311 GAGAGAGACCCCTAGCGGCG 222 8312 AGAGAGACCCCTAGCGGCGC 223 8313 GAGAGACCCCTAGCGGCGCC 224 8314 AGAGACCCCTAGCGGCGCCG 225 8315 GAGACCCCTAGCGGCGCCGC 226 8316 AGACCCCTAGCGGCGCCGCC 227 8317 GACCCCTAGCGGCGCCGCCG 228 8318 ACCCCTAGCGGCGCCGCCGG 229 8319 CCCCTAGCGGCGCCGCCGGG 230 8320 CCCTAGCGGCGCCGCCGGGG 231 8321 CCTAGCGGCGCCGCCGGGGA 232 8322 CTAGCGGCGCCGCCGGGGAA 233 8323 TAGCGGCGCCGCCGGGGAAC 234 8324 AGCGGCGCCGCCGGGGAACT 235 8325 GCGGCGCCGCCGGGGAACTG 236 8326 CGGCGCCGCCGGGGAACTGC 237 8327 GGCGCCGCCGGGGAACTGCG 238 8328 GCGCCGCCGGGGAACTGCGC 239 8329 CGCCGCCGGGGAACTGCGCC 240 8330 GCCGCCGGGGAACTGCGCCC 241 8331 CCGCCGGGGAACTGCGCCCG 242 8332 CGCCGGGGAACTGCGCCCGC 243 8333 GCCGGGGAACTGCGCCCGCT 244 8334 CCGGGGAACTGCGCCCGCTC 245 8335 CGGGGAACTGCGCCCGCTCG 246 8336 GGGGAACTGCGCCCGCTCGC 247 8337 GGGAACTGCGCCCGCTCGCG 248 8338 GGAACTGCGCCCGCTCGCGC 249 8339 GAACTGCGCCCGCTCGCGCC 250 8340 AACTGCGCCCGCTCGCGCCG 251 8341 ACTGCGCCCGCTCGCGCCGG 252 8342 CTGCGCCCGCTCGCGCCGGG 253 8343 TGCGCCCGCTCGCGCCGGGA 254 8344 GCGCCCGCTCGCGCCGGGAG 255 8345 CGCCCGCTCGCGCCGGGAGG 256 8346 GCCCGCTCGCGCCGGGAGGG 257 8347 CCCGCTCGCGCCGGGAGGGG 258 8348 CCGCTCGCGCCGGGAGGGGC 259 8349 CGCTCGCGCCGGGAGGGGCC 260 8350 GCTCGCGCCGGGAGGGGCCC 261 8351 CTCGCGCCGGGAGGGGCCCT 262 8352 TCGCGCCGGGAGGGGCCCTC 263 8353 CGCGCCGGGAGGGGCCCTCG 264 8354 GCGCCGGGAGGGGCCCTCGC 265 8355 CGCCGGGAGGGGCCCTCGCG 266 8356 GCCGGGAGGGGCCCTCGCGC 267 8357 CCGGGAGGGGCCCTCGCGCC 268 8358 CGGGAGGGGCCCTCGCGCCC 269 8359 GGGAGGGGCCCTCGCGCCCC 270 8360 GGAGGGGCCCTCGCGCCCCG 271 8361 GAGGGGCCCTCGCGCCCCGC 272 8362 AGGGGCCCTCGCGCCCCGCG 273 8363 GGGGCCCTCGCGCCCCGCGC 274 8364 GGGCCCTCGCGCCCCGCGCC 275 8365 GGCCCTCGCGCCCCGCGCCC 276 8366 GCCCTCGCGCCCCGCGCCCA 277 8367 CCCTCGCGCCCCGCGCCCAC 278 8368 CCTCGCGCCCCGCGCCCACA 279 8369 CTCGCGCCCCGCGCCCACAG 280 8370 TCGCGCCCCGCGCCCACAGG 281 8371 CGCGCCCCGCGCCCACAGGT 282 8372 GCGCCCCGCGCCCACAGGTG 283 8373 CGCCCCGCGCCCACAGGTGC 284 8374 GCCCCGCGCCCACAGGTGCA 285 8375 CCCCGCGCCCACAGGTGCAC 286 8376 CCCGCGCCCACAGGTGCACG 287 8377 CCGCGCCCACAGGTGCACGC 288 8378 CGCGCCCACAGGTGCACGCG 289 8379 GCGCCCACAGGTGCACGCGC 290 8380 CGCCCACAGGTGCACGCGCC 291 8381 GCCCACAGGTGCACGCGCCC 292 8382 CCCACAGGTGCACGCGCCCT 293 8383 CCACAGGTGCACGCGCCCTT 294 8384 CACAGGTGCACGCGCCCTTG 295 8385 ACAGGTGCACGCGCCCTTGG 296 8386 CAGGTGCACGCGCCCTTGGC 297 8387 AGGTGCACGCGCCCTTGGCG 298 8388 GGTGCACGCGCCCTTGGCGC 299 8389 GTGCACGCGCCCTTGGCGCC 300 8390 TGCACGCGCCCTTGGCGCCG 301 8391 GCACGCGCCCTTGGCGCCGC 302 8392 CACGCGCCCTTGGCGCCGCC 303 8393 ACGCGCCCTTGGCGCCGCCT 304 8394 CGCGCCCTTGGCGCCGCCTG 305 8395 GCGCCCTTGGCGCCGCCTGC 306 8396 CGCCCTTGGCGCCGCCTGCA 307 8397 GCCCTTGGCGCCGCCTGCAC 308 8398 CCCTTGGCGCCGCCTGCACC 309 8399 CCTTGGCGCCGCCTGCACCC 310 8400 CTTGGCGCCGCCTGCACCCC 311 8401 TTGGCGCCGCCTGCACCCCA 312 8402 TGGCGCCGCCTGCACCCCAC 313 8403 GGCGCCGCCTGCACCCCACG 314 8404 GCGCCGCCTGCACCCCACGC 315 8405 CGCCGCCTGCACCCCACGCG 316 8406 GCCGCCTGCACCCCACGCGC 317 8407 CCGCCTGCACCCCACGCGCC 318 8408 CGCCTGCACCCCACGCGCCC 319 8409 GCCTGCACCCCACGCGCCCC 320 8410 CCTGCACCCCACGCGCCCCC 321 8411 CTGCACCCCACGCGCCCCCT 322 8412 TGCACCCCACGCGCCCCCTC 323 8413 GCACCCCACGCGCCCCCTCC 324 8414 CACCCCACGCGCCCCCTCCG 325 8415 ACCCCACGCGCCCCCTCCGC 326 8416 CCCCACGCGCCCCCTCCGCT 327 8417 CCCACGCGCCCCCTCCGCTC 328 8418 CCACGCGCCCCCTCCGCTCC 329 8419 CACGCGCCCCCTCCGCTCCC 330 8420 ACGCGCCCCCTCCGCTCCCC 331 8421 CGCGCCCCCTCCGCTCCCCG 332 8422 GCGCCCCCTCCGCTCCCCGG 333 8423 CGCCCCCTCCGCTCCCCGGC 334 8424 GCCCCCTCCGCTCCCCGGCC 335 8425 CCGAGAGAGACCCCTAGCGG 220 8426 CCCGAGAGAGACCCCTAGCG 219 8427 ACCCGAGAGAGACCCCTAGC 218 8428 CACCCGAGAGAGACCCCTAG 217 8429 GCACCCGAGAGAGACCCCTA 216 8430 GGCACCCGAGAGAGACCCCT 215 8431 CGGCACCCGAGAGAGACCCC 214 8432 TCGGCACCCGAGAGAGACCC 213 8433 CTCGGCACCCGAGAGAGACC 212 8434 GCTCGGCACCCGAGAGAGAC 211 8435 CGCTCGGCACCCGAGAGAGA 210 8436 CCGCTCGGCACCCGAGAGAG 209 8437 CCCGCTCGGCACCCGAGAGA 208 8438 CCCCGCTCGGCACCCGAGAG 207 8439 ACCCCGCTCGGCACCCGAGA 206 8440 CACCCCGCTCGGCACCCGAG 205 8441 CCACCCCGCTCGGCACCCGA 204 8442 CCCACCCCGCTCGGCACCCG 203 8443 GCCCACCCCGCTCGGCACCC 202 8444 GGCCCACCCCGCTCGGCACC 201 8445 CGGCCCACCCCGCTCGGCAC 200 8446 CCGGCCCACCCCGCTCGGCA 199 8447 TCCGGCCCACCCCGCTCGGC 198 8448 ATCCGGCCCACCCCGCTCGG 197 8449 GATCCGGCCCACCCCGCTCG 196 8450 TGATCCGGCCCACCCCGCTC 195 8451 CTGATCCGGCCCACCCCGCT 194 8452 GCTGATCCGGCCCACCCCGC 193 8453 AGCTGATCCGGCCCACCCCG 192 8454 CAGCTGATCCGGCCCACCCC 191 8455 TCAGCTGATCCGGCCCACCC 190 8456 GTCAGCTGATCCGGCCCACC 189 8457 AGTCAGCTGATCCGGCCCAC 188 8458 GAGTCAGCTGATCCGGCCCA 187 8459 CGAGTCAGCTGATCCGGCCC 186 8460 GCGAGTCAGCTGATCCGGCC 185 8461 GGCGAGTCAGCTGATCCGGC 184 8462 AGGCGAGTCAGCTGATCCGG 183 8463 CAGGCGAGTCAGCTGATCCG 182 8464 CCAGGCGAGTCAGCTGATCC 181 8465 GCCAGGCGAGTCAGCTGATC 180 8466 AGCCAGGCGAGTCAGCTGAT 179 8467 GAGCCAGGCGAGTCAGCTGA 178 8468 AGAGCCAGGCGAGTCAGCTG 177 8469 CAGAGCCAGGCGAGTCAGCT 176 8470 TCAGAGCCAGGCGAGTCAGC 175 8471 CTCAGAGCCAGGCGAGTCAG 174 8472 GCTCAGAGCCAGGCGAGTCA 173 8473 GGCTCAGAGCCAGGCGAGTC 172 8474 GGGCTCAGAGCCAGGCGAGT 171 8475 CGCCCGCTCGCGCCGGGAGGGGCCCTCG 256 8476 GCCCGCTCGCGCCGGGAGGG 257 8477 CCCGCTCGCGCCGGGAGGGG 258 8478 CCGCTCGCGCCGGGAGGGGC 259 8479 CGCTCGCGCCGGGAGGGGCC 260 8480 GCTCGCGCCGGGAGGGGCCC 261 8481 CTCGCGCCGGGAGGGGCCCT 262 8482 TCGCGCCGGGAGGGGCCCTC 263 8483 CGCGCCGGGAGGGGCCCTCG 264 8484 GCGCCGGGAGGGGCCCTCGC 265 8485 CGCCGGGAGGGGCCCTCGCG 266 8486 GCCGGGAGGGGCCCTCGCGC 267 8487 CCGGGAGGGGCCCTCGCGCC 268 8488 CGGGAGGGGCCCTCGCGCCC 269 8489 GGGAGGGGCCCTCGCGCCCC 270 8490 GGAGGGGCCCTCGCGCCCCG 271 8491 GAGGGGCCCTCGCGCCCCGC 272 8492 AGGGGCCCTCGCGCCCCGCG 273 8493 GGGGCCCTCGCGCCCCGCGC 274 8494 GGGCCCTCGCGCCCCGCGCC 275 8495 GGCCCTCGCGCCCCGCGCCC 276 8496 GCCCTCGCGCCCCGCGCCCA 277 8497 CCCTCGCGCCCCGCGCCCAC 278 8498 CCTCGCGCCCCGCGCCCACA 279 8499 CTCGCGCCCCGCGCCCACAG 280 8500 TCGCGCCCCGCGCCCACAGG 281 8501 CGCGCCCCGCGCCCACAGGT 282 8502 GCGCCCCGCGCCCACAGGTG 283 8503 CGCCCCGCGCCCACAGGTGC 284 8504 GCCCCGCGCCCACAGGTGCA 285 8505 CCCCGCGCCCACAGGTGCAC 286 8506 CCCGCGCCCACAGGTGCACG 287 8507 CCGCGCCCACAGGTGCACGC 288 8508 CGCGCCCACAGGTGCACGCG 289 8509 GCGCCCACAGGTGCACGCGC 290 8510 CGCCCACAGGTGCACGCGCC 291 8511 GCCCACAGGTGCACGCGCCC 292 8512 CCCACAGGTGCACGCGCCCT 293 8513 CCACAGGTGCACGCGCCCTT 294 8514 CACAGGTGCACGCGCCCTTG 295 8515 ACAGGTGCACGCGCCCTTGG 296 8516 CAGGTGCACGCGCCCTTGGC 297 8517 AGGTGCACGCGCCCTTGGCG 298 8518 GGTGCACGCGCCCTTGGCGC 299 8519 GTGCACGCGCCCTTGGCGCC 300 8520 TGCACGCGCCCTTGGCGCCG 301 8521 GCACGCGCCCTTGGCGCCGC 302 8522 CACGCGCCCTTGGCGCCGCC 303 8523 ACGCGCCCTTGGCGCCGCCT 304 8524 CGCGCCCTTGGCGCCGCCTG 305 8525 GCGCCCTTGGCGCCGCCTGC 306 8526 CGCCCTTGGCGCCGCCTGCA 307 8527 GCCCTTGGCGCCGCCTGCAC 308 8528 CCCTTGGCGCCGCCTGCACC 309 8529 CCTTGGCGCCGCCTGCACCC 310 8530 CTTGGCGCCGCCTGCACCCC 311 8531 TTGGCGCCGCCTGCACCCCA 312 8532 TGGCGCCGCCTGCACCCCAC 313 8533 GGCGCCGCCTGCACCCCACG 314 8534 GCGCCGCCTGCACCCCACGC 315 8535 CGCCGCCTGCACCCCACGCG 316 8536 GCCGCCTGCACCCCACGCGC 317 8537 CCGCCTGCACCCCACGCGCC 318 8538 CGCCTGCACCCCACGCGCCC 319 8539 GCCTGCACCCCACGCGCCCC 320 8540 CCTGCACCCCACGCGCCCCC 321 8541 CTGCACCCCACGCGCCCCCT 322 8542 TGCACCCCACGCGCCCCCTC 323 8543 GCACCCCACGCGCCCCCTCC 324 8544 CACCCCACGCGCCCCCTCCG 325 8545 ACCCCACGCGCCCCCTCCGC 326 8546 CCCCACGCGCCCCCTCCGCT 327 8547 CCCACGCGCCCCCTCCGCTC 328 8548 CCACGCGCCCCCTCCGCTCC 329 8549 CACGCGCCCCCTCCGCTCCC 330 8550 ACGCGCCCCCTCCGCTCCCC 331 8551 CGCGCCCCCTCCGCTCCCCG 332 8552 GCGCCCCCTCCGCTCCCCGG 333 8553 CGCCCCCTCCGCTCCCCGGC 334 8554 GCCCCCTCCGCTCCCCGGCC 335 8555 GCGCCCGCTCGCGCCGGGAG 255 8556 TGCGCCCGCTCGCGCCGGGA 254 8557 CTGCGCCCGCTCGCGCCGGG 253 8558 ACTGCGCCCGCTCGCGCCGG 252 8559 AACTGCGCCCGCTCGCGCCG 251 8560 GAACTGCGCCCGCTCGCGCC 250 8561 GGAACTGCGCCCGCTCGCGC 249 8562 GGGAACTGCGCCCGCTCGCG 248 8563 GGGGAACTGCGCCCGCTCGC 247 8564 CGGGGAACTGCGCCCGCTCG 246 8565 CCGGGGAACTGCGCCCGCTC 245 8566 GCCGGGGAACTGCGCCCGCT 244 8567 CGCCGGGGAACTGCGCCCGC 243 8568 CCGCCGGGGAACTGCGCCCG 242 8569 GCCGCCGGGGAACTGCGCCC 241 8570 CGCCGCCGGGGAACTGCGCC 240 8571 GCGCCGCCGGGGAACTGCGC 239 8572 GGCGCCGCCGGGGAACTGCG 238 8573 CGGCGCCGCCGGGGAACTGC 237 8574 GCGGCGCCGCCGGGGAACTG 236 8575 AGCGGCGCCGCCGGGGAACT 235 8576 TAGCGGCGCCGCCGGGGAAC 234 8577 CTAGCGGCGCCGCCGGGGAA 233 8578 CCTAGCGGCGCCGCCGGGGA 232 8579 CCCTAGCGGCGCCGCCGGGG 231 8580 CCCCTAGCGGCGCCGCCGGG 230 8581 ACCCCTAGCGGCGCCGCCGG 229 8582 GACCCCTAGCGGCGCCGCCG 228 8583 AGACCCCTAGCGGCGCCGCC 227 8584 GAGACCCCTAGCGGCGCCGC 226 8585 AGAGACCCCTAGCGGCGCCG 225 8586 GAGAGACCCCTAGCGGCGCC 224 8587 AGAGAGACCCCTAGCGGCGC 223 8588 GAGAGAGACCCCTAGCGGCG 222 8589 CGAGAGAGACCCCTAGCGGC 221 8590 CCGAGAGAGACCCCTAGCGG 220 8591 CCCGAGAGAGACCCCTAGCG 219 8592 ACCCGAGAGAGACCCCTAGC 218 8593 CACCCGAGAGAGACCCCTAG 217 8594 GCACCCGAGAGAGACCCCTA 216 8595 GGCACCCGAGAGAGACCCCT 215 8596 CGGCACCCGAGAGAGACCCC 214 8597 TCGGCACCCGAGAGAGACCC 213 8598 CTCGGCACCCGAGAGAGACC 212 8599 GCTCGGCACCCGAGAGAGAC 211 8600 CGCTCGGCACCCGAGAGAGA 210 8601 CCGCTCGGCACCCGAGAGAG 209 8602 CCCGCTCGGCACCCGAGAGA 208 8603 CCCCGCTCGGCACCCGAGAG 207 8604 ACCCCGCTCGGCACCCGAGA 206 8605 CACCCCGCTCGGCACCCGAG 205 8606 CCACCCCGCTCGGCACCCGA 204 8607 CCCACCCCGCTCGGCACCCG 203 8608 GCCCACCCCGCTCGGCACCC 202 8609 GGCCCACCCCGCTCGGCACC 201 8610 CGGCCCACCCCGCTCGGCAC 200 8611 CCGGCCCACCCCGCTCGGCA 199 8612 TCCGGCCCACCCCGCTCGGC 198 8613 ATCCGGCCCACCCCGCTCGG 197 8614 GATCCGGCCCACCCCGCTCG 196 8615 TGATCCGGCCCACCCCGCTC 195 8616 CTGATCCGGCCCACCCCGCT 194 8617 GCTGATCCGGCCCACCCCGC 193 8618 AGCTGATCCGGCCCACCCCG 192 8619 CAGCTGATCCGGCCCACCCC 191 8620 TCAGCTGATCCGGCCCACCC 190 8621 GTCAGCTGATCCGGCCCACC 189 8622 AGTCAGCTGATCCGGCCCAC 188 8623 GAGTCAGCTGATCCGGCCCA 187 8624 CGAGTCAGCTGATCCGGCCC 186 8625 GCGAGTCAGCTGATCCGGCC 185 8626 GGCGAGTCAGCTGATCCGGC 184 8627 AGGCGAGTCAGCTGATCCGG 183 8628 CAGGCGAGTCAGCTGATCCG 182 8629 CCAGGCGAGTCAGCTGATCC 181 8630 GCCAGGCGAGTCAGCTGATC 180 8631 AGCCAGGCGAGTCAGCTGAT 179 8632 GAGCCAGGCGAGTCAGCTGA 178 8633 AGAGCCAGGCGAGTCAGCTG 177 8634 CAGAGCCAGGCGAGTCAGCT 176 8635 TCAGAGCCAGGCGAGTCAGC 175 8636 CTCAGAGCCAGGCGAGTCAG 174 8637 GCTCAGAGCCAGGCGAGTCA 173 8638 GGCTCAGAGCCAGGCGAGTC 172 8639 GGGCTCAGAGCCAGGCGAGT 171 8640 CGCGCCCACAGGTGCACGCGCCCTTGGCG 289 8641 GCGCCCACAGGTGCACGCGC 290 8642 CGCCCACAGGTGCACGCGCC 291 8643 GCCCACAGGTGCACGCGCCC 292 8644 CCCACAGGTGCACGCGCCCT 293 8645 CCACAGGTGCACGCGCCCTT 294 8646 CACAGGTGCACGCGCCCTTG 295 8647 ACAGGTGCACGCGCCCTTGG 296 8648 CAGGTGCACGCGCCCTTGGC 297 8649 AGGTGCACGCGCCCTTGGCG 298 8650 GGTGCACGCGCCCTTGGCGC 299 8651 GTGCACGCGCCCTTGGCGCC 300 8652 TGCACGCGCCCTTGGCGCCG 301 8653 GCACGCGCCCTTGGCGCCGC 302 8654 CACGCGCCCTTGGCGCCGCC 303 8655 ACGCGCCCTTGGCGCCGCCT 304 8656 CGCGCCCTTGGCGCCGCCTG 305 8657 GCGCCCTTGGCGCCGCCTGC 306 8658 CGCCCTTGGCGCCGCCTGCA 307 8659 GCCCTTGGCGCCGCCTGCAC 308 8660 CCCTTGGCGCCGCCTGCACC 309 8661 CCTTGGCGCCGCCTGCACCC 310 8662 CTTGGCGCCGCCTGCACCCC 311 8663 TTGGCGCCGCCTGCACCCCA 312 8664 TGGCGCCGCCTGCACCCCAC 313 8665 GGCGCCGCCTGCACCCCACG 314 8666 GCGCCGCCTGCACCCCACGC 315 8667 CGCCGCCTGCACCCCACGCG 316 8668 GCCGCCTGCACCCCACGCGC 317 8669 CCGCCTGCACCCCACGCGCC 318 8670 CGCCTGCACCCCACGCGCCC 319 8671 GCCTGCACCCCACGCGCCCC 320 8672 CCTGCACCCCACGCGCCCCC 321 8673 CTGCACCCCACGCGCCCCCT 322 8674 TGCACCCCACGCGCCCCCTC 323 8675 GCACCCCACGCGCCCCCTCC 324 8676 CACCCCACGCGCCCCCTCCG 325 8677 ACCCCACGCGCCCCCTCCGC 326 8678 CCCCACGCGCCCCCTCCGCT 327 8679 CCCACGCGCCCCCTCCGCTC 328 8680 CCACGCGCCCCCTCCGCTCC 329 8681 CACGCGCCCCCTCCGCTCCC 330 8682 ACGCGCCCCCTCCGCTCCCC 331 8683 CGCGCCCCCTCCGCTCCCCG 332 8684 GCGCCCCCTCCGCTCCCCGG 333 8685 CGCCCCCTCCGCTCCCCGGC 334 8686 GCCCCCTCCGCTCCCCGGCC 335 8687 CCGCGCCCACAGGTGCACGC 288 8688 CCCGCGCCCACAGGTGCACG 287 8689 CCCCGCGCCCACAGGTGCAC 286 8690 GCCCCGCGCCCACAGGTGCA 285 8691 CGCCCCGCGCCCACAGGTGC 284 8692 GCGCCCCGCGCCCACAGGTG 283 8693 CGCGCCCCGCGCCCACAGGT 282 8694 TCGCGCCCCGCGCCCACAGG 281 8695 CTCGCGCCCCGCGCCCACAG 280 8696 CCTCGCGCCCCGCGCCCACA 279 8697 CCCTCGCGCCCCGCGCCCAC 278 8698 GCCCTCGCGCCCCGCGCCCA 277 8699 GGCCCTCGCGCCCCGCGCCC 276 8700 GGGCCCTCGCGCCCCGCGCC 275 8701 GGGGCCCTCGCGCCCCGCGC 274 8702 AGGGGCCCTCGCGCCCCGCG 273 8703 GAGGGGCCCTCGCGCCCCGC 272 8704 GGAGGGGCCCTCGCGCCCCG 271 8705 GGGAGGGGCCCTCGCGCCCC 270 8706 CGGGAGGGGCCCTCGCGCCC 269 8707 CCGGGAGGGGCCCTCGCGCC 268 8708 GCCGGGAGGGGCCCTCGCGC 267 8709 CGCCGGGAGGGGCCCTCGCG 266 8710 GCGCCGGGAGGGGCCCTCGC 265 8711 CGCGCCGGGAGGGGCCCTCG 264 8712 TCGCGCCGGGAGGGGCCCTC 263 8713 CTCGCGCCGGGAGGGGCCCT 262 8714 GCTCGCGCCGGGAGGGGCCC 261 8715 CGCTCGCGCCGGGAGGGGCC 260 8716 CCGCTCGCGCCGGGAGGGGC 259 8717 CCCGCTCGCGCCGGGAGGGG 258 8718 GCCCGCTCGCGCCGGGAGGG 257 8719 CGCCCGCTCGCGCCGGGAGG 256 8720 GCGCCCGCTCGCGCCGGGAG 255 8721 TGCGCCCGCTCGCGCCGGGA 254 8722 CTGCGCCCGCTCGCGCCGGG 253 8723 ACTGCGCCCGCTCGCGCCGG 252 8724 AACTGCGCCCGCTCGCGCCG 251 8725 GAACTGCGCCCGCTCGCGCC 250 8726 GGAACTGCGCCCGCTCGCGC 249 8727 GGGAACTGCGCCCGCTCGCG 248 8728 GGGGAACTGCGCCCGCTCGC 247 8729 CGGGGAACTGCGCCCGCTCG 246 8730 CCGGGGAACTGCGCCCGCTC 245 8731 GCCGGGGAACTGCGCCCGCT 244 8732 CGCCGGGGAACTGCGCCCGC 243 8733 CCGCCGGGGAACTGCGCCCG 242 8734 GCCGCCGGGGAACTGCGCCC 241 8735 CGCCGCCGGGGAACTGCGCC 240 8736 GCGCCGCCGGGGAACTGCGC 239 8737 GGCGCCGCCGGGGAACTGCG 238 8738 CGGCGCCGCCGGGGAACTGC 237 8739 GCGGCGCCGCCGGGGAACTG 236 8740 AGCGGCGCCGCCGGGGAACT 235 8741 TAGCGGCGCCGCCGGGGAAC 234 8742 CTAGCGGCGCCGCCGGGGAA 233 8743 CCTAGCGGCGCCGCCGGGGA 232 8744 CCCTAGCGGCGCCGCCGGGG 231 8745 CCCCTAGCGGCGCCGCCGGG 230 8746 ACCCCTAGCGGCGCCGCCGG 229 8747 GACCCCTAGCGGCGCCGCCG 228 8748 AGACCCCTAGCGGCGCCGCC 227 8749 GAGACCCCTAGCGGCGCCGC 226 8750 AGAGACCCCTAGCGGCGCCG 225 8751 GAGAGACCCCTAGCGGCGCC 224 8752 AGAGAGACCCCTAGCGGCGC 223 8753 GAGAGAGACCCCTAGCGGCG 222 8754 CGAGAGAGACCCCTAGCGGC 221 8755 CCGAGAGAGACCCCTAGCGG 220 8756 CCCGAGAGAGACCCCTAGCG 219 8757 ACCCGAGAGAGACCCCTAGC 218 8758 CACCCGAGAGAGACCCCTAG 217 8759 GCACCCGAGAGAGACCCCTA 216 8760 GGCACCCGAGAGAGACCCCT 215 8761 CGGCACCCGAGAGAGACCCC 214 8762 TCGGCACCCGAGAGAGACCC 213 8763 CTCGGCACCCGAGAGAGACC 212 8764 GCTCGGCACCCGAGAGAGAC 211 8765 CGCTCGGCACCCGAGAGAGA 210 8766 CCGCTCGGCACCCGAGAGAG 209 8767 CCCGCTCGGCACCCGAGAGA 208 8768 CCCCGCTCGGCACCCGAGAG 207 8769 ACCCCGCTCGGCACCCGAGA 206 8770 CACCCCGCTCGGCACCCGAG 205 8771 CCACCCCGCTCGGCACCCGA 204 8772 CCCACCCCGCTCGGCACCCG 203 8773 GCCCACCCCGCTCGGCACCC 202 8774 GGCCCACCCCGCTCGGCACC 201 8775 CGGCCCACCCCGCTCGGCAC 200 8776 CCGGCCCACCCCGCTCGGCA 199 8777 TCCGGCCCACCCCGCTCGGC 198 8778 ATCCGGCCCACCCCGCTCGG 197 8779 GATCCGGCCCACCCCGCTCG 196 8780 TGATCCGGCCCACCCCGCTC 195 8781 CTGATCCGGCCCACCCCGCT 194 8782 GCTGATCCGGCCCACCCCGC 193 8783 AGCTGATCCGGCCCACCCCG 192 8784 CAGCTGATCCGGCCCACCCC 191 8785 TCAGCTGATCCGGCCCACCC 190 8786 GTCAGCTGATCCGGCCCACC 189 8787 AGTCAGCTGATCCGGCCCAC 188 8788 GAGTCAGCTGATCCGGCCCA 187 8789 CGAGTCAGCTGATCCGGCCC 186 8790 GCGAGTCAGCTGATCCGGCC 185 8791 GGCGAGTCAGCTGATCCGGC 184 8792 AGGCGAGTCAGCTGATCCGG 183 8793 CAGGCGAGTCAGCTGATCCG 182 8794 CCAGGCGAGTCAGCTGATCC 181 8795 GCCAGGCGAGTCAGCTGATC 180 8796 AGCCAGGCGAGTCAGCTGAT 179 8797 GAGCCAGGCGAGTCAGCTGA 178 8798 AGAGCCAGGCGAGTCAGCTG 177 8799 CAGAGCCAGGCGAGTCAGCT 176 8800 TCAGAGCCAGGCGAGTCAGC 175 8801 CTCAGAGCCAGGCGAGTCAG 174 8802 GCTCAGAGCCAGGCGAGTCA 173 8803 GGCTCAGAGCCAGGCGAGTC 172 8804 GGGCTCAGAGCCAGGCGAGT 171 8805 GGCCGACGGCCCACCTGGGCTTCG 351 8806 GCCGACGGCCCACCTGGGCT 352 8807 CCGACGGCCCACCTGGGCTT 353 8808 CGACGGCCCACCTGGGCTTC 354 8809 GACGGCCCACCTGGGCTTCG 355 8810 ACGGCCCACCTGGGCTTCGT 356 8811 CGGCCCACCTGGGCTTCGTG 357 8812 GGCCCACCTGGGCTTCGTGA 358 8813 GCCCACCTGGGCTTCGTGAA 359 8814 CCCACCTGGGCTTCGTGAAC 360 8815 CCACCTGGGCTTCGTGAACA 361 8816 CACCTGGGCTTCGTGAACAG 362 8817 ACCTGGGCTTCGTGAACAGT 363 8818 CCTGGGCTTCGTGAACAGTG 364 8819 CTGGGCTTCGTGAACAGTGG 365 8820 TGGGCTTCGTGAACAGTGGG 366 8821 GGGCTTCGTGAACAGTGGGA 367 8822 GGCTTCGTGAACAGTGGGAG 368 8823 GCTTCGTGAACAGTGGGAGG 369 8824 CTTCGTGAACAGTGGGAGGG 370 8825 CGCTGAGGCTCTAGAAAAGTCGAGAG 446 8826 ACGCTGAGGCTCTAGAAAAG 445 8827 GACGCTGAGGCTCTAGAAAA 444 8828 GGACGCTGAGGCTCTAGAAA 443 8829 AGGACGCTGAGGCTCTAGAA 442 8830 TAGGACGCTGAGGCTCTAGA 441 8831 CTAGGACGCTGAGGCTCTAG 440 8832 CCTAGGACGCTGAGGCTCTA 439 8833 TCCTAGGACGCTGAGGCTCT 438 8834 GTCCTAGGACGCTGAGGCTC 437 8835 AGTCCTAGGACGCTGAGGCT 436 8836 GAGTCCTAGGACGCTGAGGC 435 8837 TGAGTCCTAGGACGCTGAGG 434 8838 GTGAGTCCTAGGACGCTGAG 433 8839 GGTGAGTCCTAGGACGCTGA 432 8840 AGGTGAGTCCTAGGACGCTG 431 8841 AAGGTGAGTCCTAGGACGCT 430 8842 AAAGGTGAGTCCTAGGACGC 429 8843 CTCGTCCCCGTGAGCTTGAATCATCCGACCC 480 8844 TCGTCCCCGTGAGCTTGAAT 481 8845 CGTCCCCGTGAGCTTGAATC 482 8846 GTCCCCGTGAGCTTGAATCA 483 8847 TCCCCGTGAGCTTGAATCAT 484 8848 CCCCGTGAGCTTGAATCATC 485 8849 CCCGTGAGCTTGAATCATCC 486 8850 CCGTGAGCTTGAATCATCCG 487 8851 CGTGAGCTTGAATCATCCGA 488 8852 GTGAGCTTGAATCATCCGAC 489 8853 TGAGCTTGAATCATCCGACC 490 8854 GAGCTTGAATCATCCGACCC 491 8855 AGCTTGAATCATCCGACCCC 492 8856 GCTTGAATCATCCGACCCCG 493 8857 CTTGAATCATCCGACCCCGC 494 8858 TTGAATCATCCGACCCCGCA 495 8859 TGAATCATCCGACCCCGCAG 496 8860 GAATCATCCGACCCCGCAGG 497 8861 AATCATCCGACCCCGCAGGC 498 8862 ATCATCCGACCCCGCAGGCC 499 8863 TCATCCGACCCCGCAGGCCT 500 8864 CATCCGACCCCGCAGGCCTC 501 8865 ATCCGACCCCGCAGGCCTCC 502 8866 TCCGACCCCGCAGGCCTCCC 503 8867 CCGACCCCGCAGGCCTCCCG 504 8868 CGACCCCGCAGGCCTCCCGG 505 8869 GACCCCGCAGGCCTCCCGGG 506 8870 ACCCCGCAGGCCTCCCGGGG 507 8871 CCCCGCAGGCCTCCCGGGGG 508 8872 CCCGCAGGCCTCCCGGGGGT 509 8873 CCGCAGGCCTCCCGGGGGTG 510 8874 CGCAGGCCTCCCGGGGGTGT 511 8875 GCAGGCCTCCCGGGGGTGTC 512 8876 CAGGCCTCCCGGGGGTGTCG 513 8877 AGGCCTCCCGGGGGTGTCGT 514 8878 GGCCTCCCGGGGGTGTCGTA 515 8879 GCCTCCCGGGGGTGTCGTAT 516 8880 CCTCCCGGGGGTGTCGTATA 517 8881 CTCCCGGGGGTGTCGTATAA 518 8882 TCCCGGGGGTGTCGTATAAA 519 8883 CCCGGGGGTGTCGTATAAAG 520 8884 CCGGGGGTGTCGTATAAAGG 521 8885 GCTCGTCCCCGTGAGCTTGA 479 8886 TGCTCGTCCCCGTGAGCTTG 478 8887 CTGCTCGTCCCCGTGAGCTT 477 8888 CCTGCTCGTCCCCGTGAGCT 476 8889 TCCTGCTCGTCCCCGTGAGC 475 8890 CTCCTGCTCGTCCCCGTGAG 474 8891 GCTCCTGCTCGTCCCCGTGA 473 8892 CGCTCCTGCTCGTCCCCGTG 472 8893 GCGCTCCTGCTCGTCCCCGT 471 8894 AGCGCTCCTGCTCGTCCCCG 470 8895 GAGCGCTCCTGCTCGTCCCC 469 8896 AGAGCGCTCCTGCTCGTCCC 468 8897 GAGAGCGCTCCTGCTCGTCC 467 8898 CGAGAGCGCTCCTGCTCGTC 466 8899 TCGAGAGCGCTCCTGCTCGT 465 8900 GTCGAGAGCGCTCCTGCTCG 464 8901 AGTCGAGAGCGCTCCTGCTC 463 8902 AAGTCGAGAGCGCTCCTGCT 462 8903 AAAGTCGAGAGCGCTCCTGC 461 8904 AAAAGTCGAGAGCGCTCCTG 460 8905 GAAAAGTCGAGAGCGCTCCT 459 8906 AGAAAAGTCGAGAGCGCTCC 458 8907 TAGAAAAGTCGAGAGCGCTC 457 8908 CTAGAAAAGTCGAGAGCGCT 456 8909 TCTAGAAAAGTCGAGAGCGC 455 8910 CTCTAGAAAAGTCGAGAGCG 454 8911 GCTCTAGAAAAGTCGAGAGC 453 8912 AGGCGTTTCTGGAAGAGAATGAGAACG 604 8913 GGCGTTTCTGGAAGAGAATG 605 8914 GCGTTTCTGGAAGAGAATGA 606 8915 CGTTTCTGGAAGAGAATGAG 607 8916 CAGGCGTTTCTGGAAGAGAA 603 8917 GCAGGCGTTTCTGGAAGAGA 602 8918 GGCAGGCGTTTCTGGAAGAG 601 8919 GGGCAGGCGTTTCTGGAAGA 600 8920 GGGGCAGGCGTTTCTGGAAG 599 8921 TGGGGCAGGCGTTTCTGGAA 598 8922 GTGGGGCAGGCGTTTCTGGA 597 8923 GGTGGGGCAGGCGTTTCTGG 596 8924 AGGTGGGGCAGGCGTTTCTG 595 8925 GAGGTGGGGCAGGCGTTTCT 594 8926 AGAGGTGGGGCAGGCGTTTC 593 8927 CGTCAAAAGCAGGCACGAGCAACCTG 701 8928 GAACGAACCAAAGGAGCAAGGCG 742 8929 CGCTGACAAGGGTGCCTAGGCCCGG 1318 8930 GCGCTGACAAGGGTGCCTAG 1317 8931 TGCGCTGACAAGGGTGCCTA 1316 8932 TTGCGCTGACAAGGGTGCCT 1315 8933 ATTGCGCTGACAAGGGTGCC 1314 8934 CATTGCGCTGACAAGGGTGC 1313 8935 TCATTGCGCTGACAAGGGTG 1312 8936 CTCATTGCGCTGACAAGGGT 1311 8937 GCTCATTGCGCTGACAAGGG 1310 8938 TGCTCATTGCGCTGACAAGG 1309 8939 TTGCTCATTGCGCTGACAAG 1308 8940 CTTGCTCATTGCGCTGACAA 1307 8941 CCTTGCTCATTGCGCTGACA 1306 8942 CCCTTGCTCATTGCGCTGAC 1305 8943 TCCCTTGCTCATTGCGCTGA 1304 8944 CTCCCTTGCTCATTGCGCTG 1303 8945 TCTCCCTTGCTCATTGCGCT 1302 8946 CTCTCCCTTGCTCATTGCGC 1301 8947 TCTCTCCCTTGCTCATTGCG 1300 8948 CGCAATTCCGTATTTGTTCCGG 1738 8949 GCAATTCCGTATTTGTTCCG 1739 8950 CAATTCCGTATTTGTTCCGG 1740 8951 AATTCCGTATTTGTTCCGGG 1741 8952 ATTCCGTATTTGTTCCGGGT 1742 8953 TTCCGTATTTGTTCCGGGTC 1743 8954 TCCGTATTTGTTCCGGGTCT 1744 8955 CCGTATTTGTTCCGGGTCTG 1745 8956 CGTATTTGTTCCGGGTCTGC 1746 8957 GTATTTGTTCCGGGTCTGCA 1747 8958 TATTTGTTCCGGGTCTGCAT 1748 8959 ATTTGTTCCGGGTCTGCATG 1749 8960 TTTGTTCCGGGTCTGCATGA 1750 8961 TTGTTCCGGGTCTGCATGAG 1751 8962 TGTTCCGGGTCTGCATGAGC 1752 8963 GTTCCGGGTCTGCATGAGCA 1753 8964 TTCCGGGTCTGCATGAGCAA 1754 8965 TCCGGGTCTGCATGAGCAAA 1755 8966 CCGGGTCTGCATGAGCAAAT 1756 8967 CGGGTCTGCATGAGCAAATA 1757 8968 CCGCAATTCCGTATTTGTTC 1737 8969 GTACGTTGGCAGACGCAGTGACG 4923 8970 TACGTTGGCAGACGCAGTGA 4924 8971 ACGTTGGCAGACGCAGTGAC 4925 8972 CGTTGGCAGACGCAGTGACG 4926 8973 GTTGGCAGACGCAGTGACGT 4927 8974 TTGGCAGACGCAGTGACGTA 4928 8975 TGGCAGACGCAGTGACGTAT 4929 8976 GGCAGACGCAGTGACGTATT 4930 8977 GCAGACGCAGTGACGTATTT 4931 8978 CAGACGCAGTGACGTATTTG 4932 8979 AGACGCAGTGACGTATTTGA 4933 8980 GACGCAGTGACGTATTTGAG 4934 8981 ACGCAGTGACGTATTTGAGA 4935 8982 CGCAGTGACGTATTTGAGAG 4936 8983 GCAGTGACGTATTTGAGAGT 4937 8984 TGTACGTTGGCAGACGCAGT 4922 8985 ATGTACGTTGGCAGACGCAG 4921 8986 CATGTACGTTGGCAGACGCA 4920 8987 TCATGTACGTTGGCAGACGC 4919 8988 ATCATGTACGTTGGCAGACG 4918 8989 TATCATGTACGTTGGCAGAC 4917 8990 GTATCATGTACGTTGGCAGA 4916 8991 GGTATCATGTACGTTGGCAG 4915 8992 GGGTATCATGTACGTTGGCA 4914 8993 TGGGTATCATGTACGTTGGC 4913 8994 CTGGGTATCATGTACGTTGG 4912 8995 GCTGGGTATCATGTACGTTG 4911 8996 TGCTGGGTATCATGTACGTT 4910 8997 TTGCTGGGTATCATGTACGT 4909 8998 CTTGCTGGGTATCATGTACG 4908

Hot Zones (Relative upstream location to gene start site)   1-800 1200-1800 4800-5100

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11978) TGAAGACTATAGCCCCTTTCTTTTGGCTGATTTCTCCCTTTTGGAATGGG AATGTTTACCCAGTGCTGGTACTCCCATTATATCTTGGAAGTAAATAACT TGTTTTGATTTTACAGGCTCATAGATGGAAAGAGATGAGTCTCAGATGAG ACTTTGGACTTGGAATTTGGACTTTGGACTTTTGAGTTAATGCTGGAACA AGTCAAGACCTTGGAGGACTGTTGGGAAGGCATGATTGTATTTTGAAATG AGAGAGGGACATGAGATTTGAGAGGGGCTGGGGCAGAATGATATAGTTTG TGTACTTATCCCCACCCAAATCTCATGTGGAATTGTAATCCCTAGTATTG GAGGCGGGACCTGGTAGGAGGTGATTGGATCATGGGGGTGGATTACTCAT GAATCGTTTAGCACTATCTGTTTGTTGCTGTCCTTGTGATGAGTGACTTC TCATGAGATCTGGCTGTTTAAAAGTGTGTGGCACCATGCTTTCTCTTGTT CCCGGTCTGGTCATGTGACATCCCTACTCCCCCTTCACCTTCCATCATGA TTGTAAGTTTCCTGAGGCCTCACCAGAAGCCAAGCAGATGCCAGCATCAG GCTTCCTGCAAAGCCTGAAGAATCATAAGCTGATTAAATCTCTCTCTCTT TTTTTTTGAGATGGAGTCTCATCCTGTGGCCCACGCTGGAAGGCAGTGGC ACTATCTCTGCTCTGAGGCTCAAGTGATTCTCCTGCCTCAGCCTCCTGAG TAGCTAGGATTACAGGTGCATGCCACCACACCCAGCTAATTTTTGTATTT TTAGTAGAGATGAGGTTTCACCACGTTGGCCAGGCTGGTCTCGAACTCCT GACCTAAGGTTATCTGTCCACCTCAGCTTCCCAAAGTGCTGGGATTACAG GTGTGAGCCATTGCACCCGGCCTCTTTTCTTTATAAACTACCTAGTCTCA GATATTTCTTTTTGGCAATGCAAGAATGGCCTAATACACCAAGATTTGTG TTTTCACCATTTTTTTTCTGTTTATAACCATGTTATTTTACTGTTTATCT GAGAAAAGAACCTGGCCTGTTAGTATTTTTTATCGACTGACAAACTCACC AGAATAAAGTGGGTATTAGGACCCTTGGTTCTGCAAGATTTTGGTAACAG AATCTGTTTTCACTAATTGTTGGGAAACAGAAATGATATTCCTTTTAGAC CTAGCTCCCTAAACCTTTTCCTCGTTTTGCTTTTTGGTACAATAATGAGG GCTGGCAGGGCTACTTGACACCATTAGCAGTAGACAAATTTTTCAATAAG GACTAACAGAGAAAAACTATGGAAATTCTGATTTATTGTTTGGCCGAGAG AGTTCTCTGTCTCCTTGTGCCCTTGCTCTCCATGTATATTTTATGAGACA TTTCAGCAAAGGCATCTCTCAAAATTAATCCCATAGCTGTCTCCTTCAAT TCTCCTCCTTGAAACTCTTCACCAATCTTCATAGGGCCTTGGCCACTTCT TGAGGGCCCTCCCACCAGATGATTAAAGGCAATACAGTGAACGAAAGTCT TATTCCGAGACTTGTCTTTGTAAACTTAGTGATCCTTCTTCCTTTTCACT TACGAAAATTTAAAGAGAAGCAGTCTCAAATGTGAACTGAATGCCGTCCC ATTACTCCCCCAAACTGGGAAGAAGCTGGTCATATACTTGCACATTTATA TAATAAATATTCAAAGACTCTAATCTGGTATCTTCCATATAACACACACT TTAACTCCTATTTTAAACTTTCAAAAGGCTTTTTATGGCATCTTATGCCC TACTTTAAAATGTCTGTCAGCCTAATATTTCTACTTTTTTTTTATTAATT TATTTTCAAGGTCATGTGTGAAAACAACTTTCAGTGAAAAGAACCCATCT GCTTTGACAAAAATGTACACTATAAACCTTCACTTCTACAAGGGTCTAAA AAAATTCAAGGGTTTGATTCGAATGCTTCCAAACCACATTCCCTAAAGCA TGCTTTGTGCAACACTAGGAGTTGGTGCAGCAAACAAAAACAAGGAATGG GGAGAGGTGCCAGGCAGGCGCGGTGGCTCATGCCTGTAATCCCAGCACTT TGGGTGACCCAGGCAGGTGGATCACTTGAGGTCAGGAGTTCGAGATCAGT CAACATGGTGAAACCCCGTCTCTTCTAAAATACAAAAATTAGCTAGGTGT GTGTGCACGTAATCCCAGCACTTTGTGAGGCTGAGGTGGGTGGATCACTT GAGGTCAGGAGTTCAAGACCAGCCTGGCCAACATGGTGAAACCCCATCTC TACTAAAAATACAAAAAAAAAAATTAGCCGGGTATGGTGGCACACACCTG TAATCCCAGCCACTCAGGAGGCTGAGGCATGAGAATTGCTTGATCCCTGG AGGCAGAGGTTGCAGTGAGCAGAGATCATGCCTGGGGAAAAGAGAGAGAC TATGTCTCAAAAAAAAAAAAAAGAAAAAAGAAATAGGGAGAGGGAGTGAT GCTACATACTGAAGCTACCTACCCTCTTTAAAATTCAAAATGCAGATTAT CATCTTAAAGAATTGTATGCATTTTAAAGTGAAGGTATTTGTTTAATTTT GTTTAACCTCTTATTTCTCAAAGTTACTTGATTACAGAATTCTTTTCGGG TATGCATGCAATACCGAGCAACATATGACCGAGCTAGGGTTCCATGGTAC ACAGTTTGGAACTTGTGCTAGAAAATGTTACTCATTATTTTATAAGAAGT ATTATTCCAAGGCATCTATGCATTGGAAAATAATCTTGATATGAAATATG ACTCGATCCCTTCTCATACCATATTTACAGGGTATGGTGGTTAACTTTAT GTGTCCACTTGTCTCTTAGCCCATCCTGGGTTAAGGGATGCCCAGACAGC TGGTAAAATATTATTTCTGGGAGAGTCTGTGAGAGTATTTCTGGAAGAGA TTAGCATTTGAATCAATAGACTGGGTATCAAAAACCCATCCTCACTAATG TGAGTGGGCATCATTCAATTAACTGAGCATAGGAATCATAAAAGAGGTGG AGGAAGGCAAATTTGCTCTCTCTTCTGAAGCTGGGACATCCATCATCTGC TCCTGCCCTTGACTTCAGAGCTCCAGGTTCTTGAGCCTTCAATACCAGCA GACACTCTTCAGCCCTTCAGCTTCACATGCAAGAATTACACTGGATTTCC TGGTTCTTCGGCTTGCAGGGGGCACATATTGGAACTTCTTGGCCTTTATA ATCTCCTGAGCCAATTCCTATAAGAAGTGTCTTTATATATCTATATATAT CCCATTGGTTCTGTTTGTTTGGAAAACCCTGATACACAGGGCTATCTATA GCTCACCCCCCAAGTACTAAGTCTCCAGATGATTATTAGGTTCTTATAAA CACAAAATATATATGCTTATTTTGTAATATCAGGTTGTTAACTTCATCTG AGTAGTTTTCAGCACATATGATGGGGATAAGTCATCAGTTATGACAGACA ACTTCCGGAGTTACATCACCAATGTTCATCTATCACTCACCTTTGCTTCA AACTTTATGCATCATATTCAAGTTAATTCCTATTGCATGTACTTCTGATT TCCACCTACAAAATGAACATTAAATTTATTATTTCCCTCATTTAAAAAAA AAGATTTCAGGCCAGGTGCAGTGGCTCATGCCTGCAATCACAGCACTTTG GGAGGCTGAGGCGGGTGGATCACTTGAGGCCAGGAGTTCAAGACCAGCCT AGGCAACATGATGAAACCCCTATCTCTACTAAAAATACAAAATACAAAAA AAAAAAAAATTAGCCAGGTGTATGTGTGCCTGCAATCCCACCCCTACTCA GGAGGCTGAGGCAGGAGGATAGCTTGAGTTCTGGAGGCGGAGGTTGCCGT GAGCCAAGATCACGCCACTGTACTCAAGCCTGGGCAAGACAGCAAGACTG AGACTCTGTCTCAAAAAAAAAAAAAAAAAAAGTTTTCAAATCACTTTTTC TCCTTTGCAGTTCAGCATTGTCACTAAAGGTGTCCAGGTTAGACAGAACT GGGTACAAACCCTCCTTTCTCTTCCCCTTACTCTTTCACCTCTTCATTTG TGACATGCAATGTTAACCCAATACAACTCATGAGTATTCAGAGATCCCGT CTGTATACGTCCTCAGCCTGACATACTGTAATCCTTAGGCATCCTTATTA GTAATAAGATGTCCTTGTGTGATTTTTTCACAAACTTTTCACAGACCCTA TCTATGTTCATTCCTGGAACCTCTGGCACATTCTTCTTCCTTCTCTTCCC AATCTCAACTTTTTCATCCTCTGAATCTCCCTATACTTTCCCCGTGGACA AGCCTCTAGAAATGTTAAAATGTCAGATCATGATTGGTAAATCTGTAGTG ACTAATTGCCCACTGCTGCCTATTCCATCTGACCTAAATTCCTCAGGTCT TCTAACATTAAGACCTCTTCCTGGCCGGTTGTGCTGGCTCATGCCTGTAA TTCCAACACTTTAGGCAGCTGAGGCAGGCAGATCACTTGAGGTCAAGAGT TCAAGACCAGCCTGGCCAACATGGTGAAGCCCTGTCTGTACTAAAAAATA CAAAAATTAGCCGGACCTGGTGGTGCGTGCCTGTAATCCCAGCTACTCGG GAGGCTGAGTCAGGAGAATCACTTGAACCCGGGGCAGCGGGGGAGGCTGC AGTGAGTGGAGATCAAACCACCGCACTCCAGCCCAGGTGACAGAGCAAGA GTCAGTCTCAAAAAAAAAAAAACAAAAAAAAAAACCTCTTCCTATAGCTA ACTCCCACTTACCACCCCCATCATGAACACTCTTGATGTATTTACATGGT TTCTCCTTCGAACATCCTCCTTTCTTCTTTCTTAATGGTTGTTATCAAAT ACCCTGATAAAAAACAAAAACAAAAAACCTCCTCTGAAGGTCCCTTATTC ACCCTTCCAACGCTACAGGTCTGTAACTCTCATTTTCTTTTTAAAAAATT TTTATTTTTTTAATTTATTTTATTTTTTTTTTCAGACGGAGTCTTGCTCT GTCGCCCAGGCTGGAGTGCAGTGACACGATCTCGGCTCACTGCAACCTCC ACTTCCCAGGTTCAAGCAATTCTCCTGCCTCAGCCTCCTGAGTTGCTGGG ATTACAGGTGCCTACCACCACACCTGACCTCAAGTAACCCACCCACCTCG ACCTCCCAAAGTGCTGAGAATACAGGTGTGAGCCATCATGCCTGGCCAAA ATTTTTAAATTTTAAAAAATATATTTTATTTTTTGTAGAGACAGGGTCTC ATTTTGAGCCCAAACTGGTCTTGAACTCCTAGGCTCAAGTGATCCTCCTG CCTTGGCCTCCCAAAATGCTGGGATTATAGGCACAAGCCACCAGGCCTGA TCCTTACTTTTCTTCTGATGAATTCACATATATGTGCACAAATACTTTAT ACTAAATTGTATTTACTGATGTACTTTTTTCACTGTGCCTTTTCTTTTTC TTGCCCAGATATTTTTCTCATATAAACATTAGCTCCTTAATGGGAGCAAA TGAACCAGTTTTTTTTTAATTCCCACCCAAAGTGAGAATATAAAAATTTT TTATTGATCCACCAATACTGAACACTTTCATTTCTAATAGTTATATTTAA CTGAATAAATTACACACGGGACAAAAATGTTATTTAAGGGATAAAGTTGG GTGTTTGCTCAGGGACAACGTTGTATATTGAATGATTTGGTGCTTTTGTG AATTTATCATTCAAAAGACCATCGTGATGGCTAAATAACAGAAAGGAGAG CTTTATTGGCAATATCAATTTGCAAACCCGGAAGACATAGTCTTCGGTGT ATGCTGAATGTGGTCTCTCTTCAAAAGAGAGGAAGGACAGTTGGGTTTCA TGCCTCACAGGGTCTGTTTCACACAGTGGAGTCATACATATTCAGCAGGT TTGGAGGAAAAGATATACATATTTATGAGGGGAGCTGAGTGCATGTGCAA TGGGTAAATATGTATGTGACATCCCATGTACACTTTGGGGCAGGGTTTTA GTGTTAAAATGAGGTAAAATTTGGCTCTTTACATCAAAAGGTGAACTACA GGACCCAAAGACAGTTTGTGCACAGCCTCTAATAAACTGGCTGACACTGG CTTAAGGTCTGCAATTGCTTATCAGAAAAGAATGTTTGTAAGGCTGGTCC TCATTCCAATTAGAGTTGTAGTGGTCTGGGTTGTAAATCACAGGATGGGG CTGATAGTTCCTATTATTAGGGAGTTTAGAGCCATAGAAATTGAGAAATT GGTCATGCCAGCCAGTCCCCGAACCCTAACCCTGTAGGTAACTTTGTTTC CTTAACCTTACAGTCCATCTTAGGTGATAAAGGGGTGTCTGTTTTGGTAT CTCACATCACAAATTGTTGGTTGGTTTGTGTGTTTGTTTCATCATTCAGG ATGTTGTTTCTTTAGGGAATGTGAACCTGAATTCTCAAGGCTTGTTAGAC TGTAATGTTCCCATTCATTTTAGGTTTAGCTCATGCTTCTCTAGCCACAG CCTTCACTTGGATTTTAAAAGTTGAATTACTCATCAAAGTCTCTAGGACA CGAAAGACAATCCTTAGGTATGATTTGACCAGTAAAAAAGAGATCCAGCT GCCTTGAAGCATAAGATCCCCTCGGCTCCAATGTCTATCACTAATATTCA GTGTGGCAAGGATCCCAGGCCACAGAGCTGTGGCTTCCTGCAGCTGCTCT GGGGAGTGACTCTCTTGGAGCATGTGATGTGGTCTTCCATTGTGCAGGAC CAGCCCAGTGGCATCCTTTCAACACCTCTGGCAAGCAGCCTTTCCAAGCA CGGGTGCCGTCTGAAAACAGGAGGCATATCTTTCACATCCTAGGCACACG CCCTAGGGAGTGGTCAGGGTTTTGTCCAGTTCTCAGCAAACTAGCTACAG CTCCATCCCTTACTCCCACACTCAAGAGAGATACTAGAATACAACTGAGA GTAGCCTGATATGATGCTAACCTCGAGTTGCTTTTATTTAAATTAAAATA AATCAACCAGACACAGTGGCTCATGCCTATAATCCCTGCATTTTAGGAGA TCAAGGAGGGTGGATCACGAGCTCAGGAGTTTAAGACCAACCTGGATAAC ATGGCAAGACCCCATCTCTACAAAAAGTACACAAATTAGCTGGACATGGT AGTGCGCACCTGTAGCCCCAGCTACTCTGGAGGCTGAGGTGAAGGATCAC TTGAGCCCAGGAGGTAGAGGTTGCAGTGAGCTGAGATTGTGCCACTGCTA ATAATTAATTAAATAATTAATTAAATTAATAAATCGTGCCACTTTATTAA ATAAATAAAACAAGAGTAAATCACTCACAAATTTGGAGCTTTTATTAGCA AAACATTACTTAGGAAATCTAAATAAATAACACGGGGTTGACAGCCATTG TTCTAACTGGCAGCCCCTGGCAAGCTCAAAGCCAGGATTATGCTGGTCAC TTAAGTGACAGCTATTGCGAATTGTTGTTCTCTCAAGAAAAAAGAACCGA TTTCTATGGTAAACCAGGCACTGTGCTGGGTGCCTTTACAATTCATCACC ACACCACCTAATGAAAGGAGCATTCTTCAGAAACTGTAGTGCTCAGGCTT TCTCAAGGCCTGAGTTCTTTTCCACCAGAGCATATTGTTGCCCTATTATC CAAAGTTCTCTAAGGAAGAGAACTGACGTAAGACCCACATGGCTCCATTA CATCTTCTGGCTACTTGATTGATTTTCATACTCCCTACCTCTGGGGTTGG TATGTACTATCTATTTCTTTCTCCTCTCGTTCTTCCTTTTTATTCCATAA AATACAGGAATATTCCTGTACATTAGTCCTTGCAGCAACCTTGGAATTAC TACATTCCTCAAACAAGTTATGGAAGCCAGCTGCCAATATTGGTCCCTGG TTAAACAGTGAATTCTGTTGTTCCATAGAGTTACTACTGAAATACCTAAG CCATTTTGTAAAATATAATTTAGTTGATCTGAAGGCTGTCTCTAAAGCAG TTTTATGTAGTGATTACAGAGAAGGACTAATTTCAAGAGTATTTTATTGT TTAAAAAAATGTAAACATTTTATGGATGCACTAGTGAAGTAAAGACCAAT AAATGAAGCAGTAACTTTAATAAAAGGGTAAGTAAAATGTCACATCCTCT GCCTATATTCAGGTCTGTTAGGTATGTGTAGTTAAATGTAGGTAAGTTAG TTGATAATTATTTATTTAAGCATTTCTTTATGTCTACTCATTAAAAAGAA AAAAAGATTAAAAGAATGTTACTATGTGAAAAACTGCCCATCACTGGGGA AAAGAATTTTATTATGCAAAGCTTCAACGCTATTTACAGTTTAGACTTTT GTAGCTATTGAAGGCTGACATTGAGATAAAGAAGTTAATCATGTCCTTCT GTCTTGGAGGAGGTAGAAAGAGATGAGAATGAATACAATTCAGGATCTAC TTCTGGTCTTTGATGAGGAGTTAGCACACGGTTCTGGGAGGAAAGACAGG TTAAGAGGCATGTGAAACTCTCAAATACGTCACTGCGTCTGCCAACGTAC ATGATACCCAGCAAGCTCACATCTTCATGGAAAGCATGGTAATTCCCAAC ACTACCGGAAGTCTGGAGTGGCTAAGTAATCCATATATTCAACCAGGAAG CAGCTAAAGAAATATTCTAATTACCTAGGAAGGTTTCTGATTTCAAAAGG ACATGAATAAAAAGTAGAAGGAATCCACTCCCAAGGACGGACATCAGAGT AGCTTAAAATGTGAGAATAATTTTAGGGGAATTTTAGAGGTTTGGTTATA GACTTATGTTCCCCCAAAATTCATATGTTGAAGCCCTAACCCCCAGTACC TTAGAACATGACTGTATTTGGGTAGGGCCTTTGAAGAGCTAATTAAATTA AGGCCACTGGCGTGGGCCCTAATATAATCTGGCTGGTATTCTTGTAAGAG GAGGAGATTAGGACACACAGAAATACCAGAGGTACCTGTGCAGAGGAAAG AACGTGTGAGGACTTAGCAAGGGTGCAGCCATCTGCAAGCCAAGGAGACC TCTGAGGATTCCAATCCTATCTGCATCTTGATCTTAGACTTTTCTGGAAC TGTGAGAAAATAAATTTCTTGGTTTAAGCCACCCAGTCTGTGATATTTTG TTATGGCAGCTCTAGTAAACTAATACAGATTTTAAATGTCATTAAATGTC AATGTTTAAGCTTTGACAAAATTTTCTAAAGGAAAGTATAAAAGGTCATT TTCTTTCTTTTCAGAGCCTGATGATTGCGGGAGGGGTAAGCCAGCTGCAT GGGGATCATGATGCAATGCTGATGCAGGACAGACAGAAAGTAGATCTCTT CCATTTCTATTTTTTTTTTTTCTGTTGAGTTGAATGATCTTCAGACTGAA AATGAAAGAAAGGTCACTGGAAATAAAGGCCAAAGATGAGTGACAGGATT ATAGAATAAGTCTTAGCTGTTCTAAAGAAGGACATATTATGTACCCCCAC CCCCAAATTCATATGTTGAAGTCCTAACCCGACAGTGTCTCAAAATGTGA CCATATTTGGAGATAGGGTCAAAGATGTAATTAAGGTTAAATGAGGTCAT TAGCATGGATCCTAACCCAATATCTGCTGTCCTTATAACAAGAGGAGATT AGGGCACAGTAAGACACAGAGGGAAGACCATGTGAGAATACAGGGAGAAG GTGGCCATCTGCAAGCCAAGGAGAGAGGCCTCAGAAGTAACCAACTCAGC CAACACCTCGATTTCAGACTTCCAGCCTCCTGAAATGTGAGGAAATACAT TTCTGGTGTTTGATCCATCCAGTCTATGGTAAGTTATGGCACCCCTGCAG GGTTCATCTGGCTCAGACTTAACGATTGCTTTTGGTGATATTTATAGGGC ACAGATAACAGCCTAAACACAAGACGACAGAAACGCGGCCCAGCAGACTA TGCATAAAATAGAAATGGGGTATCTGGACCAATTGGAGTCTGCAGTGGGA TGCGGTTACTAAAACAGTCAAATGCAACATGAGGCTCCAGGCAGAGTAGT GGGCAACATCTCCCATGTTGCAGCAGTCAGAGCACACTTCGAGTACTGTA AAAAGACACAGACAAGCCAGAACACATTTAGAGAATGGCCAAGGTGTGGA AGGAACCAGAAACCATGCCATTATGCAACTGTTGAAGGAAGTGCCTGTTT TACCTTGTGAAGAGAAGACTCTAGAGGAAGAAGTAGCATGAAAACCGCTG GCAAATTTGTAAAGATCTGAAGTGTGGAAAAGAATTATTCTGCTTGGTCA CTGGGGATACAAGGATATCTGAGTGGGAGTTTAAAGGCGGGGGATGTGAG CTTTAAATGGGATAAGAACATTCTAGTAACCAGAAATGCCCAAAGATAGA ATGCACAGTCTGGAGAGCCAGTGAATATCTCACAAATGGAGACACTTGAA ACTAGGATGGGGATGCTGTTGTAGGAATTCCAGCAGACAAGTGGTTGTTG GTTCCTTCCCCAACTTTGTAGGGTTATAACTAGGGATGTTCCTGCGTTTT CTGCTTGGAGGATCTGCAAGACACCTCAGGGCAGGAAATGGCATTAAATG CAGAACAGAGCTAGTGGCTGAAAAGCAAAAAGCCATCAGGATCTCTGAGT AGTGAAGGAACCAGAGAACATGCAGGCAATGTCCATCATTCTGACGCAAT CAGCAGCATAATCATCTTCCCCCAGGAACATCTTGACCAGGGAATGTGTC AGTGTGGTGAATTTCAACAGTGGAAAGAGAAACTGCTAAATCTAAGAACT TTAATTTTTATAGATTATGATCTCATCTCTACAATTTTGAATTTCATGCT CAATAAAAGTTCCTTACTCTCTTTTTTTTTTTTTGAGACGGAGTCTCGCT CTGTCGCCCAGGCTGGAGTGCAGTGGCGCGATCTCGGCTCACTTCAAGCT CAGCCTCCCGGGTTCACGCCATTCTCCTGCCTCAGCCTCCCCAGTAGCTG GGACTACAGGCGCCCGCCACGACGCCCGGCTAATTTTTTGTATTTTTAGT AGAGACGGGGTTTCACCGTGTTAGCCAGGATGGTGTTGATCTCCTGACCT CGTGATCCGCCCGCCTCAGCCTCCCAAAGAAAAGTCCCTCACTCTTAAAG TTGCCTCCTCCTTCCCAGGGCTGGCTTCATGGGCATGCAACCCTGGAGAG TCTCACAGGCCCTGCGGTGGGAGGAGCCCCATGCTTGGTTTAACGCTCTG CCATTGCCATCTTAAAATTCTTAATTTAATTTTTTTTCTTTTTTTTGAGG TGGAGTCTCGCTCTGTCGCCCAGGCTGGAGTGCAATGGCACAATCTTGGC TCACTGCAACCTCCGCCTCCCAGGTTCAAGCGATTCTCCTGCCTCAGCCT CTGGAGTAGCTGGGATTACAGGCAGGAGTAACCACGCTCGGCTAATTTTT GCATTTTTAGTAGAGATGGGGGTTTCACCATGTTGGCCAGGCTGGTCTAG AACTCCTGACCTCAGGTGATCTGCCCACCTGGGCCTCCTAAAGTGCTGGG ATTACAGGCATGAGCCACCAGGCCCGGCCTTAAAATTCTTAATAATGTAA CAAAGGGTCTCACGTTTGCATTTTGCAGTGGACTCTGCAAGATTTGTAGC TTTGGACCACGTTTCTCTTTGCATTCAGATACCTTCTTTTTTGCCTTATT TGCTCATGCAGACCCGGAACAAATACGGAATTGCGGTGGGTAAATGTGGT GCAGAAAGTGAACAACTGGGTTTGTCCTGTCACTTTAGGCTTTTCCCTGC TGTCCCAGCTTCATGTCACTTACTTGCTATTAGATTTGGGAGTTCATTAG CTTCATTTTCCTGATGTATAAATAGGAATAATAGTAACAGCCTCTTTGGC TTTTGTAGGAAGTAAATGACATGAAGCGTATAAACAAATACTGCATGACA ATAAATATTTGTCCTTATTTGTTGAGGACATCCAAAGGACATTCAGGGGC AAAAGTAATCCAAGAGTCAAGACTGAATGCCTAGTGCGGGAAAAGACACA CAAGACAACATTTAGGGGAGCTGGTACAGAAATGACTTCCCAGGAAGGAA GTCTGTACCCCGCTGGCTGAGCCATCCTTCCCGGGCCTAGGCACCCTTGT CAGCGCAATGAGCAAGGGAGAGAAGGCAGGCTGCAGTGCAGCCCTCAGAA GGGCCAGAGCACTCCCTGGCTTCAGTCCTTCGCTCCAAGCCCTGTGTGGA GTGGGCTGTGGCTTGGTAACTAAATGCTACTTCAGGTCAAGAGCAGGGGA TATATCTGGGCAGTTCTAGAGCATTCTAAACTATCTGGACACTAACTGGA CAGTGGACGGTTTGTGTTTAATCCAGGAGAAAGTGGCATGGCAGAAGGTT CATTTCTATAATTCAGGACAGACACAATGAAGAACAAGGGCAGCGTTTGA GGTCAGAAGTCCTCATTTACGGGGGTCGAATACGAATGATCTCTCCTAAT TTTTCCTTCTTCCCCAACTCAGATGGATGTTACATCCCTGCTTAACAACA AAAAAAGACCCCCCGCCCCGCAAAATCCACACTGACCACCCCCTTTAACA AAACAAAACCAAAAACAAACAAAAATATAAGAAAGAAACAAAACCCAAGC CCAGAACCCTGCTTTCAAGAAGAAGTAAATGGGTTGGCCGCTTCTTTGCC AGGTCCTGCGCCTTGCTCCTTTGGTTCGTTCTAAAGATAGAAATTCCAGG TTGCTCGTGCCTGCTTTTGACGTTGGGGGTTAAAAAATGAGGTTTTGCTG TCTCAACAAGCAAAGAAAATCCTATTTCCTTTAAGCTTCACTCGTTCTCA TTCTCTTCCAGAAACGCCTGCCCCACCTCTCCAAACCGAGAGAAAAAACG AAATGCGGATAAAAACGCACCCTAGCAGCAGTCCTTTATACGACACCCCC GGGAGGCCTGCGGGGTCGGATGATTCAAGCTCACGGGGACGAGCAGGAGC GCTCTCGACTTTTCTAGAGCCTCAGCGTCCTAGGACTCACCTTTCCCTGA TCCTGCACCGTCCCTCTCCTGGCCCCAGACTCTCCCTCCCACTGTTCACG AAGCCCAGGTGGGCCGTCGGCCGGGGAGCGGAGGGGGCGCGTGGGGTGCA GGCGGCGCCAAGGGCGCGTGCACCTGTGGGCGCGGGGCGCGAGGGCCCCT CCCGGCGCGAGCGGGCGCAGTTCCCCGGCGGCGCCGCTAGGGGTCTCTCT CGGGTGCCGAGCGGGGTGGGCCGGATCAGCTGACTCGCCTGGCTCTGAGC CCCGCCGCCGCGCTCGGGCTCCGTCAGTTTCCTCGGCAGCGGTAGGCGAG AGCACGCGGAGGAGCGTGCGCGGGGGCCCCGGGAGACGGCGGCGGTGGCG GCGCGGGCAGAGCAAGGACGCGGCGGATCCCACTCGCACAGCAGCGCACT CGGTGCCCCGCGCAGGGTCGCG ATG

32. HAMP

Hepcidin is a peptide hormone produced by the liver. Hepcidin plays a role in maintaining iron balance by inhibiting iron absorption across the gut mucosa and transport of iron from macrophages which serve as a depot of iron storage and transport. Hepcidin production in the liver increases when iron enters liver cells from the blood thereby causing its release into the blood. In contrast, in states of high hepcidin (e.g. inflammation), serum iron levels drop because iron remains trapped in macrophages, resulting in anemia (Ganz T. 2003. Blood 102 (3): 783-8). Beta-thalassemia a common congenital anemia is characterized by excessive iron absorption and overload of iron associated with low levels hepcidin levels. In this situation, increasing expression of hepcidin may be therapeutic to treat the abnormal iron absorption in individuals with β-thalassemia and related disorders. Mutations in this gene cause hemochromatosis type 2B, also known as juvenile hemochromatosis, a disease caused by severe iron overload resulting in cardiomyopathy, cirrhosis, and endocrine failure.

Protein: HAMP Gene: HAMP (Homo sapiens, chromosome 19, 35773410-35776064 [NCBI Reference Sequence: NC_(—)000019.9]; start site location: 35773482; strand: positive)

Gene Identification GeneID 57817 HGNC 15598 HPRD 05925 MIM 606464

Targeted Sequences Relative upstream Sequence Design location to gene start ID No: ID Sequence (5′-3′) site 8999 CGTGCCGTCTGTCTGGCTGTCCCAC 1 9005 CGAGTGACAGTCGCTTTTATGGGGC 60 9035 CGGGGCATGGCCAGCAGCCGCCAGG 424 9086 CGTGTGCCCGATCCGCACGTGGTGT 563 9121 CGACAGGCTGACGGGCCAAGCTTGG 2344 9150 CGGATGGGCAGGGAGGATACCGTTT 3109 9151 CGTGGGCGGCGGCGGCTGCGTGGTG 3287

Target Shift Sequences Relative upstream Sequence location to gene ID No: Sequence (5′-3′) start site 8999 CGTGCCGTCTGTCTGGCTGTCCCAC 1 9000 GTGCCGTCTGTCTGGCTGTC 2 9001 TGCCGTCTGTCTGGCTGTCC 3 9002 GCCGTCTGTCTGGCTGTCCC 4 9003 CCGTCTGTCTGGCTGTCCCA 5 9004 CGTCTGTCTGGCTGTCCCAC 6 9005 CGAGTGACAGTCGCTTTTATGGGGC 60 9006 GAGTGACAGTCGCTTTTATG 61 9007 AGTGACAGTCGCTTTTATGG 62 9008 GTGACAGTCGCTTTTATGGG 63 9009 TGACAGTCGCTTTTATGGGG 64 9010 GACAGTCGCTTTTATGGGGC 65 9011 ACAGTCGCTTTTATGGGGCC 66 9012 CAGTCGCTTTTATGGGGCCT 67 9013 AGTCGCTTTTATGGGGCCTG 68 9014 GTCGCTTTTATGGGGCCTGC 69 9015 TCGCTTTTATGGGGCCTGCC 70 9016 CGCTTTTATGGGGCCTGCCA 71 9017 CCGAGTGACAGTCGCTTTTA 59 9018 ACCGAGTGACAGTCGCTTTT 58 9019 GACCGAGTGACAGTCGCTTT 57 9020 GGACCGAGTGACAGTCGCTT 56 9021 GGGACCGAGTGACAGTCGCT 55 9022 TGGGACCGAGTGACAGTCGC 54 9023 CTGGGACCGAGTGACAGTCG 53 9024 TCTGGGACCGAGTGACAGTC 52 9025 GTCTGGGACCGAGTGACAGT 51 9026 TGTCTGGGACCGAGTGACAG 50 9027 GTGTCTGGGACCGAGTGACA 49 9028 GGTGTCTGGGACCGAGTGAC 48 9029 TGGTGTCTGGGACCGAGTGA 47 9030 CTGGTGTCTGGGACCGAGTG 46 9031 TCTGGTGTCTGGGACCGAGT 45 9032 CTCTGGTGTCTGGGACCGAG 44 9033 GCTCTGGTGTCTGGGACCGA 43 9034 TGCTCTGGTGTCTGGGACCG 42 9035 CGGGGCATGGCCAGCAGCCGCCAGG 424 9036 GGGGCATGGCCAGCAGCCGC 425 9037 GGGCATGGCCAGCAGCCGCC 426 9038 GGCATGGCCAGCAGCCGCCA 427 9039 GCATGGCCAGCAGCCGCCAG 428 9040 CATGGCCAGCAGCCGCCAGG 429 9041 ATGGCCAGCAGCCGCCAGGC 430 9042 TGGCCAGCAGCCGCCAGGCT 431 9043 GGCCAGCAGCCGCCAGGCTC 432 9044 GCCAGCAGCCGCCAGGCTCC 433 9045 CCAGCAGCCGCCAGGCTCCT 434 9046 CAGCAGCCGCCAGGCTCCTC 435 9047 AGCAGCCGCCAGGCTCCTCA 436 9048 GCAGCCGCCAGGCTCCTCAG 437 9049 CAGCCGCCAGGCTCCTCAGG 438 9050 AGCCGCCAGGCTCCTCAGGA 439 9051 GCCGCCAGGCTCCTCAGGAG 440 9052 CCGCCAGGCTCCTCAGGAGT 441 9053 CGCCAGGCTCCTCAGGAGTG 442 9054 ACGGGGCATGGCCAGCAGCC 423 9055 CACGGGGCATGGCCAGCAGC 422 9056 ACACGGGGCATGGCCAGCAG 421 9057 CACACGGGGCATGGCCAGCA 420 9058 GCACACGGGGCATGGCCAGC 419 9059 TGCACACGGGGCATGGCCAG 418 9060 ATGCACACGGGGCATGGCCA 417 9061 CATGCACACGGGGCATGGCC 416 9062 ACATGCACACGGGGCATGGC 415 9063 TACATGCACACGGGGCATGG 414 9064 CTACATGCACACGGGGCATG 413 9065 CCTACATGCACACGGGGCAT 412 9066 GCCTACATGCACACGGGGCA 411 9067 CGCCTACATGCACACGGGGC 410 9068 TCGCCTACATGCACACGGGG 409 9069 ATCGCCTACATGCACACGGG 408 9070 CATCGCCTACATGCACACGG 407 9071 CCATCGCCTACATGCACACG 406 9072 CCCATCGCCTACATGCACAC 405 9073 CCCCATCGCCTACATGCACA 404 9074 TCCCCATCGCCTACATGCAC 403 9075 TTCCCCATCGCCTACATGCA 402 9076 CTTCCCCATCGCCTACATGC 401 9077 ACTTCCCCATCGCCTACATG 400 9078 CACTTCCCCATCGCCTACAT 399 9079 TCACTTCCCCATCGCCTACA 398 9080 CTCACTTCCCCATCGCCTAC 397 9081 ACTCACTTCCCCATCGCCTA 396 9082 CACTCACTTCCCCATCGCCT 395 9083 CCACTCACTTCCCCATCGCC 394 9084 TCCACTCACTTCCCCATCGC 393 9085 CTCCACTCACTTCCCCATCG 392 9086 CGTGTGCCCGATCCGCACGTGGTGT 563 9087 GTGTGCCCGATCCGCACGTG 564 9088 TGTGCCCGATCCGCACGTGG 565 9089 GTGCCCGATCCGCACGTGGT 566 9090 TGCCCGATCCGCACGTGGTG 567 9091 GCCCGATCCGCACGTGGTGT 568 9092 CCCGATCCGCACGTGGTGTT 569 9093 CCGATCCGCACGTGGTGTTT 570 9094 CGATCCGCACGTGGTGTTTT 571 9095 GATCCGCACGTGGTGTTTTC 572 9096 ATCCGCACGTGGTGTTTTCC 573 9097 TCCGCACGTGGTGTTTTCCC 574 9098 CCGCACGTGGTGTTTTCCCA 575 9099 CGCACGTGGTGTTTTCCCAG 576 9100 GCACGTGGTGTTTTCCCAGT 577 9101 CACGTGGTGTTTTCCCAGTG 578 9102 ACGTGGTGTTTTCCCAGTGT 579 9103 CGTGGTGTTTTCCCAGTGTC 580 9104 GCGTGTGCCCGATCCGCACG 562 9105 AGCGTGTGCCCGATCCGCAC 561 9106 CAGCGTGTGCCCGATCCGCA 560 9107 TCAGCGTGTGCCCGATCCGC 559 9108 ATCAGCGTGTGCCCGATCCG 558 9109 CATCAGCGTGTGCCCGATCC 557 9110 GCATCAGCGTGTGCCCGATC 556 9111 AGCATCAGCGTGTGCCCGAT 555 9112 AAGCATCAGCGTGTGCCCGA 554 9113 CAAGCATCAGCGTGTGCCCG 553 9114 GCAAGCATCAGCGTGTGCCC 552 9115 GGCAAGCATCAGCGTGTGCC 551 9116 GGGCAAGCATCAGCGTGTGC 550 9117 AGGGCAAGCATCAGCGTGTG 549 9118 CAGGGCAAGCATCAGCGTGT 548 9119 GCAGGGCAAGCATCAGCGTG 547 9120 AGCAGGGCAAGCATCAGCGT 546 9121 CGACAGGCTGACGGGCCAAGCTTGG 2344 9122 GACAGGCTGACGGGCCAAGC 2345 9123 ACAGGCTGACGGGCCAAGCT 2346 9124 CAGGCTGACGGGCCAAGCTT 2347 9125 AGGCTGACGGGCCAAGCTTG 2348 9126 GGCTGACGGGCCAAGCTTGG 2349 9127 GCTGACGGGCCAAGCTTGGC 2350 9128 CTGACGGGCCAAGCTTGGCG 2351 9129 TGACGGGCCAAGCTTGGCGC 2352 9130 GACGGGCCAAGCTTGGCGCC 2353 9131 ACGGGCCAAGCTTGGCGCCC 2354 9132 CGGGCCAAGCTTGGCGCCCT 2355 9133 GGGCCAAGCTTGGCGCCCTG 2356 9134 GGCCAAGCTTGGCGCCCTGG 2357 9135 GCCAAGCTTGGCGCCCTGGC 2358 9136 CCAAGCTTGGCGCCCTGGCC 2359 9137 CAAGCTTGGCGCCCTGGCCA 2360 9138 AAGCTTGGCGCCCTGGCCAT 2361 9139 AGCTTGGCGCCCTGGCCATC 2362 9140 GCTTGGCGCCCTGGCCATCT 2363 9141 CTTGGCGCCCTGGCCATCTG 2364 9142 TTGGCGCCCTGGCCATCTGC 2365 9143 TGGCGCCCTGGCCATCTGCC 2366 9144 GGCGCCCTGGCCATCTGCCC 2367 9145 GCGCCCTGGCCATCTGCCCT 2368 9146 CGCCCTGGCCATCTGCCCTC 2369 9147 GCGACAGGCTGACGGGCCAA 2343 9148 GGCGACAGGCTGACGGGCCA 2342 9149 AGGCGACAGGCTGACGGGCC 2341 9150 CGGATGGGCAGGGAGGATACCGTTT 3109 9151 CGTGGGCGGCGGCGGCTGCGTGGTG 3287 9152 GTGGGCGGCGGCGGCTGCGT 3288 9153 TGGGCGGCGGCGGCTGCGTG 3289 9154 GGGCGGCGGCGGCTGCGTGG 3290 9155 GGCGGCGGCGGCTGCGTGGT 3291 9156 GCGGCGGCGGCTGCGTGGTG 3292 9157 CGGCGGCGGCTGCGTGGTGG 3293 9158 GGCGGCGGCTGCGTGGTGGT 3294 9159 GCGGCGGCTGCGTGGTGGTG 3295 9160 CGGCGGCTGCGTGGTGGTGG 3296 9161 GGCGGCTGCGTGGTGGTGGC 3297 9162 GCGGCTGCGTGGTGGTGGCG 3298 9163 CGGCTGCGTGGTGGTGGCGG 3299 9164 GGCTGCGTGGTGGTGGCGGG 3300 9165 GCTGCGTGGTGGTGGCGGGC 3301 9166 CTGCGTGGTGGTGGCGGGCG 3302 9167 TGCGTGGTGGTGGCGGGCG 3303 9168 GCGTGGTGGTGGCGGGCG 3304 9169 CGTGGTGGTGGCGGGCG 3305 9170 GTGGTGGTGGCGGGCG 3306 9171 TGGTGGTGGCGGGCG 3307 9172 GCGTGGGCGGCGGCGGCTGC 3286 9173 GGCGTGGGCGGCGGCGGCTG 3285 9174 CGGCGTGGGCGGCGGCGGCT 3284 9175 CCGGCGTGGGCGGCGGCGGC 3283 9176 GCCGGCGTGGGCGGCGGCGG 3282 9177 GGCCGGCGTGGGCGGCGGCG 3281 9178 AGGCCGGCGTGGGCGGCGGC 3280

Hot Zones (Relative upstream location to gene start site)  1-630 3061-3321

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11979) CGCCCGCCACCACCACGCAGCCGCCGCCGCCCACGCCGGCCTCTGCTGCC CCCTTCCCCAGCCCTTAGCACAGAGAGGGACACATGCCCCTCCCCCAGCT GCGTTTTTTTATAGTAGATTTTTAACAAAAAACGGGGAGAAATAATGCAT TTCTGTGGATACAGTGCCCACCGCCCTCCTCCACTTGGAAACGGTATCCT CCCTGCCCATCCGTCTGTCTGTCGCCCTTCTCCCGGCCCTCACTAAGCCC CGGCACTTCTAGTGGTCTCACCTGGAGGCAAGAGGGAGGGGACAGAGGCC CTGCCACGTCCCGCTGCCTCCTGCTCTCTGGAGGTACTGAGACAGGGTGC TGATGGGAAGGAGGGGAGCCTTTGGGGGGCCACCCGGGGCCTGGACCTAT GCAGGGAGGCCACGTCCCACCCCACCTCTTGTTTCTGGGTCCCTGCTCCC CTTTGGGGGTGTGTGTGTGTGTTTTAATTTTCTTTATGGAAAAATTGACA AAAAAAAAATAGAGAGAGAGGTATTTAACTGCAATAAACTGGCCCCATGT GGCCCCCGCCTTGTCTGCTTGTGTGTTTGTCCATCTCAGGAGTGGGGAGG GGGCCTGGGGTCTGCAGAGCTCCACGAGGCATGGTTCTGCTGTTGTGCAC ATGGCTGTGCATGGTCCCTGCCAGCTGCACCACCCATTACCCAGTGGTTG GTTGGATGGATGGAGGAATTAAGGAATGAATGTCCCCTTTGAGGCCCTAG ACGTGCATGAGGGTGTGGGGAGCTGGGGTCAAGGACATGTCCCATGTTGG AGGAGAGGCAGGGGTCTCCGTGTCAACAGTTCCTGAAAACACAACCAGCC CCTGGCCCTGCCCTGCTGGGCCAAAGCCCTCCCCTCTGCACCAGCCAATA GTGGGGCCTGGCCTTGAGCCCCTCACCCCCAGGGAGGGCAGATGGCCAGG GCGCCAAGCTTGGCCCGTCAGCCTGTCGCCTTGCACCAAGGCTCTGGCGC CTGTGCTGTGACCCCTGCCCCTGCTGATGATGAAACCTGTCCTCAGCTGA GATGCAGCGATGCCTGGTAGGGCTGGGGGCTGCTCCTGTGTCTCCCCAGG TGAGCACACCCCTATTCACTGGGCCCTGCTTCAGCCTGCAGCACCCTTCA ACTCCCAGGAGCTGGGCTTGCCACTCTGCTCACCTTGTGGAGCTCCATCT GCCTTTCCTCCCCAATTCCCCCACTCCCTGCACTCGTCTCTTCCCACAAG AGCCCTGTCTCCTTTTCCTAGCTATTCCCATCTGAGGCCATCTTTATTCA TTTAGTTTTTAGAGACAGGGTTTCACTCTCACCCAGGCTGGGGTGCAGTG GCACACAATCACGGCTCACTGCAGCCTTGACCAACTACAGGTGCGTAGCA CCACAGCCAAGTTTTTGTATAGATGGGGTCTCGCTTTGTTACCCAGGCTG TGACAAGAGGAGCCTCCCACGTGGTGTGGATGAGGAGGCAGATGGCAGGG CCTGTGCATTTCTGTGCTTGAGTGGGCCTTGAAAGTGGTTCAGCAACCAG GAAGAAGTGTTCATTCCTCGACAACAACATCCCCGGGCTCTGGTGACTTG GCTGACACTGGATGGCCCTGGAATGAAAAAGGCAAAGAGGCAAAATGTGC AAGGGCCCATCTGGAACCAAGGTTTGTTGATCCCCTGGGCCGTGTGCACC CTGAGCTGGGCCTGGTAGTGGAAAGGAATGAAGGCACTGCAGTCAGGCAG CCTGGGTTCATCCCCCAGCTAGTGGTGTCCTAAGGAACCGGCTCCCCAAA AACATCCCTGGCTTGTAGTGCTTGCCAATTTCTGGGTGTCAAGACTCCCA CTGCTGCTGATTTCAGGATACCAGCATGATGCCACTGAATGCAGAGTTTC GAGATGTGCATGGTCTGCTATGTTGAGCCAGGTCTAGCATACCGCTGTGC CCTGCTGTGTTTTAGGGGAGATGGGGAAACCTGGTGGGTAAGAGCAAAAG CCCTGGAGTCAGGCTGTCCAGGCTAGAATCTCAGCTCTGCCTCTGGCTGA GCAAGCTTGGGCCATGCCCTGATCTCTGCCTTCAGTGCCTTTTCTGTAAA GTGAAGGAAATGAGTGTCCGACGGGGAGGAGGTTCCTAAAAGGGAGCAGG GTCTGGGGAGCCCAGGCCTCTGGGGTTGGGTGACTGAGAAGGCAGCCCCT GAATACAGAGCAGAGCTGAAGGTGGGGCAGTAAGTGCTGCTGGGAGAACA GGCAGCACAGGCTGAGTTGGTGCAGAAGTGAGTCAACATATGTGCCATCG TATAAAATGTACTCATCGGACTGTAGATGTTAGCTATTACTATTACTGCT ATTTTATGTTTTATAGACAGGGTCTCACTCTGTCACCCAGGCTGGAGTGC AGTCACACAATCATAGCTCACTGCAACCTCAGCCTCCTGGGCTTAAGCGA TCTGCCTCAGCCTCCCAAGTAGCTGGGACTACAGATGTGTGCCACCACGC CTGGCTAAATTTGTTTAAAATTTTTTTTGTAGAGATGGGGTCTCCCTATG TTGCCCAGGCTAGTCTTGAACTTCTGGGCTCAAGCGACCCTCCTGCCTTG GCCTCCCAAATTGCTGGGATTACAGGCATAAGCCACTGTGCTGGGCCATA TTACTGCTGTCATTTATGGCCAAAAGTTTGCTCAAACATTTTCCAGTTAC CAGAGCCACATCTCAAGGGTCTGACACTGGGAAAACACCACGTGCGGATC GGGCACACGCTGATGCTTGCCCTGCTCAGGGCTATCTAGTGTTCCCTGCC AGAACCTATGCACGTGTGGTGAGAGCTTAAAGCAATGGATGCTTCCCCCA ACATGCCAGACACTCCTGAGGAGCCTGGCGGCTGCTGGCCATGCCCCGTG TGCATGTAGGCGATGGGGAAGTGAGTGGAGGAGAGCGGAACCTTGATTCT GCTCATCAAACTGCTTAACCGCTGAAGCAAAAGGGGGAACTTTTTTCCCG ATCAGCAGAATGACATCGTGATGGGGAAAGGGCTCCCCAGATGGCTGGTG AGCAGTGTGTGTCTGTGACCCCGTCTGCCCCACCCCCTGAACACACCTCT GCCGGCTGAGGGTGACACAACCCTGTTCCCTGTCGCTCTGTTCCCGCTTA TCTCTCCCGCCTTTTCGGCGCCACCACCTTCTTGGAAATGAGACAGAGCA AAGGGGAGGGGGCTCAGACCACCGCCTCCCCTGGCAGGCCCCATAAAAGC GACTGTCACTCGGTCCCAGACACCAGAGCAAGCTCAAGACCCAGCAGTGG GACAGCCAGACAGACGGCACG ATG

HBV

Hepatitis B is an infectious inflammatory disease of the liver caused by the hepatitis B virus (HBV). About ⅓ of the world population is believed to be infected, including 350 million who are chronic carriers. Acutely symptoms include liver inflammation, vomiting and jaundice, while chronic hepatitis B is implicated in cirrhosis and liver cancer. HBV is a DNA virus that has a circular genome of partially double-stranded DNA (Zuckerman A. J. 1996 in Baron S, et al. Baron's Medical Microbiology (4th Ed)) with a full length strand with 3020-3320 nucleotides and a short-length strand of 1700-2800 nucleotides for the short length-strand (Kay A and Zoulim F. 2007 Virus research 127 (2): 164-176). HBV uses reverse transcription to replicate: virus gains entry into the cell by endocytosis, multiplies via RNA made by a host enzyme, then reversed transcribed into viral genomic DNA. The partially double stranded viral DNA is rendered fully double stranded when transformed into covalently closed circular DNA (cccDNA). cccDNA serves as a template for transcription of four viral mRNAs encoding viral proteins called C, X, P and S critical of virus infection and replication. HBV core protein is coded for by gene C (HBcAg); its DNA polymerase is encoded by gene P; the surface antigen (HBsAg) is encoded by the S gene. HBx protein is encoded by the X gene and is believed to drive cccDNA transcription and stimulates genes to promote cell growth associated with liver cancer and the persistence of HBV.

Hepatitis B Virus (1-3215 [NCBI Reference Sequence: NC_(—)003977]; strand: negative)

Targeted Sequences Relative upstream location to Sequence Design start ID No: ID Sequence (5′-3′) site 9179 CCGATTGGTGGAGGCAGGAGGAGG 72 9180 CGAGATTGAGATCTTCTGCGACGCGG 780 9235 CGCGGCGATTGAGACCTTCGTC 801 9290 CGTCTGCGAGGCGAGGGAGTTCTTCT 819 9345 CGATACAGAGCAGAGGCGGTGT 1200 9346 CGCGTAAAGAGAGGTGCGCCCCGTGG 1674 9360 ACGGGTCGTCCGCGGGATTCAGCGCCG 1754 9409 CGTCCCGCGCAGGATCCAGTTGG 1800 9432 CGGCTGCGAGCAAAACAAGCTGCTAG 1909 9468 CGCATGCGCCGATGGCCTATGGCCAA 1978 9496 CGCCGCAGACACATCCAGCGATA 2826 9525 GCTCCAGACCGGCTGCGA 1900 9561 CGTCCATCGCAGGATCCAGTTGG 1800 9562 CGCCGCAGACACATCCAGCGATA 2826 9591 CAAATGGCACTAGTAAACTGAG 2524 9592 GAGATTGAGATCTGCGGCGACGCGG 780 9593 CGACGCGGCGATTGAGATCTTCGTCTG 801 9594 AGGGGTCGTCCGCGGGATTCAGCGCCG 1754

Target Shift Sequences Relative upstream Sequence location ID No: Sequence (5′-3′) to start site 9179 CCGATTGGTGGAGGCAGGAGGAGG 72 9180 CGAGATTGAGATCTTCTGCGACGCGG 780 9181 GAGATTGAGATCTTCTGCGA 781 9182 AGATTGAGATCTTCTGCGAC 782 9183 GATTGAGATCTTCTGCGACG 783 9184 ATTGAGATCTTCTGCGACGC 784 9185 TTGAGATCTTCTGCGACGCG 785 9186 TGAGATCTTCTGCGACGCGG 786 9187 GAGATCTTCTGCGACGCGGC 787 9188 AGATCTTCTGCGACGCGGCG 788 9189 GATCTTCTGCGACGCGGCGA 789 9190 ATCTTCTGCGACGCGGCGAT 790 9191 TCTTCTGCGACGCGGCGATT 791 9192 CTTCTGCGACGCGGCGATTG 792 9193 TTCTGCGACGCGGCGATTGA 793 9194 TCTGCGACGCGGCGATTGAG 794 9195 CTGCGACGCGGCGATTGAGA 795 9196 TGCGACGCGGCGATTGAGAC 796 9197 GCGACGCGGCGATTGAGACC 797 9198 CGACGCGGCGATTGAGACCT 798 9199 GACGCGGCGATTGAGACCTT 799 9200 ACGCGGCGATTGAGACCTTC 800 9201 CGCGGCGATTGAGACCTTCG 801 9202 GCGGCGATTGAGACCTTCGT 802 9203 CGGCGATTGAGACCTTCGTC 803 9204 GGCGATTGAGACCTTCGTCT 804 9205 GCGATTGAGACCTTCGTCTG 805 9206 CGATTGAGACCTTCGTCTGC 806 9207 GATTGAGACCTTCGTCTGCG 807 9208 ATTGAGACCTTCGTCTGCGA 808 9209 TTGAGACCTTCGTCTGCGAG 809 9210 TGAGACCTTCGTCTGCGAGG 810 9211 GAGACCTTCGTCTGCGAGGC 811 9212 AGACCTTCGTCTGCGAGGCG 812 9213 GACCTTCGTCTGCGAGGCGA 813 9214 ACCTTCGTCTGCGAGGCGAG 814 9215 CCTTCGTCTGCGAGGCGAGG 815 9216 CTTCGTCTGCGAGGCGAGGG 816 9217 TTCGTCTGCGAGGCGAGGGA 817 9218 TCGTCTGCGAGGCGAGGGAG 818 9219 CGTCTGCGAGGCGAGGGAGT 819 9220 GTCTGCGAGGCGAGGGAGTT 820 9221 TCTGCGAGGCGAGGGAGTTC 821 9222 CTGCGAGGCGAGGGAGTTCT 822 9223 TGCGAGGCGAGGGAGTTCTT 823 9224 GCGAGGCGAGGGAGTTCTTC 824 9225 CGAGGCGAGGGAGTTCTTCT 825 9226 GAGGCGAGGGAGTTCTTCTT 826 9227 AGGCGAGGGAGTTCTTCTTC 827 9228 GGCGAGGGAGTTCTTCTTCT 828 9229 GCGAGGGAGTTCTTCTTCTA 829 9230 CGAGGGAGTTCTTCTTCTAG 830 9231 CCGAGATTGAGATCTTCTGC 779 9232 CCCGAGATTGAGATCTTCTG 778 9233 TCCCGAGATTGAGATCTTCT 777 9234 TTCCCGAGATTGAGATCTTC 776 9235 CGCGGCGATTGAGACCTTCGTC 801 9236 GCGGCGATTGAGACCTTCGT 802 9237 CGGCGATTGAGACCTTCGTC 803 9238 GGCGATTGAGACCTTCGTCT 804 9239 GCGATTGAGACCTTCGTCTG 805 9240 CGATTGAGACCTTCGTCTGC 806 9241 GATTGAGACCTTCGTCTGCG 807 9242 ATTGAGACCTTCGTCTGCGA 808 9243 TTGAGACCTTCGTCTGCGAG 809 9244 TGAGACCTTCGTCTGCGAGG 810 9245 GAGACCTTCGTCTGCGAGGC 811 9246 AGACCTTCGTCTGCGAGGCG 812 9247 GACCTTCGTCTGCGAGGCGA 813 9248 ACCTTCGTCTGCGAGGCGAG 814 9249 CCTTCGTCTGCGAGGCGAGG 815 9250 CTTCGTCTGCGAGGCGAGGG 816 9251 TTCGTCTGCGAGGCGAGGGA 817 9252 TCGTCTGCGAGGCGAGGGAG 818 9253 CGTCTGCGAGGCGAGGGAGT 819 9254 GTCTGCGAGGCGAGGGAGTT 820 9255 TCTGCGAGGCGAGGGAGTTC 821 9256 CTGCGAGGCGAGGGAGTTCT 822 9257 TGCGAGGCGAGGGAGTTCTT 823 9258 GCGAGGCGAGGGAGTTCTTC 824 9259 CGAGGCGAGGGAGTTCTTCT 825 9260 GAGGCGAGGGAGTTCTTCTT 826 9261 AGGCGAGGGAGTTCTTCTTC 827 9262 GGCGAGGGAGTTCTTCTTCT 828 9263 GCGAGGGAGTTCTTCTTCTA 829 9264 CGAGGGAGTTCTTCTTCTAG 830 9265 ACGCGGCGATTGAGACCTTC 800 9266 GACGCGGCGATTGAGACCTT 799 9267 CGACGCGGCGATTGAGACCT 798 9268 GCGACGCGGCGATTGAGACC 797 9269 TGCGACGCGGCGATTGAGAC 796 9270 CTGCGACGCGGCGATTGAGA 795 9271 TCTGCGACGCGGCGATTGAG 794 9272 TTCTGCGACGCGGCGATTGA 793 9273 CTTCTGCGACGCGGCGATTG 792 9274 TCTTCTGCGACGCGGCGATT 791 9275 ATCTTCTGCGACGCGGCGAT 790 9276 GATCTTCTGCGACGCGGCGA 789 9277 AGATCTTCTGCGACGCGGCG 788 9278 GAGATCTTCTGCGACGCGGC 787 9279 TGAGATCTTCTGCGACGCGG 786 9280 TTGAGATCTTCTGCGACGCG 785 9281 ATTGAGATCTTCTGCGACGC 784 9282 GATTGAGATCTTCTGCGACG 783 9283 AGATTGAGATCTTCTGCGAC 782 9284 GAGATTGAGATCTTCTGCGA 781 9285 CGAGATTGAGATCTTCTGCG 780 9286 CCGAGATTGAGATCTTCTGC 779 9287 CCCGAGATTGAGATCTTCTG 778 9288 TCCCGAGATTGAGATCTTCT 777 9289 TTCCCGAGATTGAGATCTTC 776 9290 CGTCTGCGAGGCGAGGGAGTTCTTCT 819 9291 GTCTGCGAGGCGAGGGAGTT 820 9292 TCTGCGAGGCGAGGGAGTTC 821 9293 CTGCGAGGCGAGGGAGTTCT 822 9294 TGCGAGGCGAGGGAGTTCTT 823 9295 GCGAGGCGAGGGAGTTCTTC 824 9296 CGAGGCGAGGGAGTTCTTCT 825 9297 GAGGCGAGGGAGTTCTTCTT 826 9298 AGGCGAGGGAGTTCTTCTTC 827 9299 GGCGAGGGAGTTCTTCTTCT 828 9300 GCGAGGGAGTTCTTCTTCTA 829 9301 CGAGGGAGTTCTTCTTCTAG 830 9302 TCGTCTGCGAGGCGAGGGAG 818 9303 TTCGTCTGCGAGGCGAGGGA 817 9304 CTTCGTCTGCGAGGCGAGGG 816 9305 CCTTCGTCTGCGAGGCGAGG 815 9306 ACCTTCGTCTGCGAGGCGAG 814 9307 GACCTTCGTCTGCGAGGCGA 813 9308 AGACCTTCGTCTGCGAGGCG 812 9309 GAGACCTTCGTCTGCGAGGC 811 9310 TGAGACCTTCGTCTGCGAGG 810 9311 TTGAGACCTTCGTCTGCGAG 809 9312 ATTGAGACCTTCGTCTGCGA 808 9313 GATTGAGACCTTCGTCTGCG 807 9314 CGATTGAGACCTTCGTCTGC 806 9315 GCGATTGAGACCTTCGTCTG 805 9316 GGCGATTGAGACCTTCGTCT 804 9317 CGGCGATTGAGACCTTCGTC 803 9318 GCGGCGATTGAGACCTTCGT 802 9319 CGCGGCGATTGAGACCTTCG 801 9320 ACGCGGCGATTGAGACCTTC 800 9321 GACGCGGCGATTGAGACCTT 799 9322 CGACGCGGCGATTGAGACCT 798 9323 GCGACGCGGCGATTGAGACC 797 9324 TGCGACGCGGCGATTGAGAC 796 9325 CTGCGACGCGGCGATTGAGA 795 9326 TCTGCGACGCGGCGATTGAG 794 9327 TTCTGCGACGCGGCGATTGA 793 9328 CTTCTGCGACGCGGCGATTG 792 9329 TCTTCTGCGACGCGGCGATT 791 9330 ATCTTCTGCGACGCGGCGAT 790 9331 GATCTTCTGCGACGCGGCGA 789 9332 AGATCTTCTGCGACGCGGCG 788 9333 GAGATCTTCTGCGACGCGGC 787 9334 TGAGATCTTCTGCGACGCGG 786 9335 TTGAGATCTTCTGCGACGCG 785 9336 ATTGAGATCTTCTGCGACGC 784 9337 GATTGAGATCTTCTGCGACG 783 9338 AGATTGAGATCTTCTGCGAC 782 9339 GAGATTGAGATCTTCTGCGA 781 9340 CGAGATTGAGATCTTCTGCG 780 9341 CCGAGATTGAGATCTTCTGC 779 9342 CCCGAGATTGAGATCTTCTG 778 9343 TCCCGAGATTGAGATCTTCT 777 9344 TTCCCGAGATTGAGATCTTC 776 9345 CGATACAGAGCAGAGGCGGTGT 1200 9346 CGCGTAAAGAGAGGTGCGCCCCGTGG 1674 9347 GCGTAAAGAGAGGTGCGCCC 1675 9348 CGTAAAGAGAGGTGCGCCCC 1676 9349 GTAAAGAGAGGTGCGCCCCG 1677 9350 TAAAGAGAGGTGCGCCCCGT 1678 9351 AAAGAGAGGTGCGCCCCGTG 1679 9352 AAGAGAGGTGCGCCCCGTGG 1680 9353 AGAGAGGTGCGCCCCGTGGT 1681 9354 GAGAGGTGCGCCCCGTGGTC 1682 9355 AGAGGTGCGCCCCGTGGTCG 1683 9356 GAGGTGCGCCCCGTGGTCGG 1684 9357 AGGTGCGCCCCGTGGTCGGC 1685 9358 GGTGCGCCCCGTGGTCGGCC 1686 9359 CCGCGTAAAGAGAGGTGCGC 1673 9360 ACGGGTCGTCCGCGGGATTCAGCGCCG 1754 9361 CGGGTCGTCCGCGGGATTCA 1755 9362 GGGTCGTCCGCGGGATTCAG 1756 9363 GGTCGTCCGCGGGATTCAGC 1757 9364 GTCGTCCGCGGGATTCAGCG 1758 9365 TCGTCCGCGGGATTCAGCGC 1759 9366 CGTCCGCGGGATTCAGCGCC 1760 9367 GTCCGCGGGATTCAGCGCCG 1761 9368 TCCGCGGGATTCAGCGCCGA 1762 9369 CCGCGGGATTCAGCGCCGAC 1763 9370 CGCGGGATTCAGCGCCGACG 1764 9371 GCGGGATTCAGCGCCGACGG 1765 9372 CGGGATTCAGCGCCGACGGG 1766 9373 GGGATTCAGCGCCGACGGGA 1767 9374 GGATTCAGCGCCGACGGGAC 1768 9375 GATTCAGCGCCGACGGGACG 1769 9376 ATTCAGCGCCGACGGGACGT 1770 9377 TTCAGCGCCGACGGGACGTA 1771 9378 TCAGCGCCGACGGGACGTAG 1772 9379 CAGCGCCGACGGGACGTAGA 1773 9380 AGCGCCGACGGGACGTAGAC 1774 9381 GCGCCGACGGGACGTAGACA 1775 9382 CGCCGACGGGACGTAGACAA 1776 9383 GACGGGTCGTCCGCGGGATT 1753 9384 AGACGGGTCGTCCGCGGGAT 1752 9385 GAGACGGGTCGTCCGCGGGA 1751 9386 CGAGACGGGTCGTCCGCGGG 1750 9387 CCGAGACGGGTCGTCCGCGG 1749 9388 CCCGAGACGGGTCGTCCGCG 1748 9389 CCCCGAGACGGGTCGTCCGC 1747 9390 GCCCCGAGACGGGTCGTCCG 1746 9391 GGCCCCGAGACGGGTCGTCC 1745 9392 CGGCCCCGAGACGGGTCGTC 1744 9393 ACGGCCCCGAGACGGGTCGT 1743 9394 AACGGCCCCGAGACGGGTCG 1742 9395 AAACGGCCCCGAGACGGGTC 1741 9396 CAAACGGCCCCGAGACGGGT 1740 9397 CCAAACGGCCCCGAGACGGG 1739 9398 CCCAAACGGCCCCGAGACGG 1738 9399 GCCCAAACGGCCCCGAGACG 1737 9400 GGCCCAAACGGCCCCGAGAC 1736 9401 AGGCCCAAACGGCCCCGAGA 1735 9402 GAGGCCCAAACGGCCCCGAG 1734 9403 AGAGGCCCAAACGGCCCCGA 1733 9404 TAGAGGCCCAAACGGCCCCG 1732 9405 GTAGAGGCCCAAACGGCCCC 1731 9406 GGTAGAGGCCCAAACGGCCC 1730 9407 CGGTAGAGGCCCAAACGGCC 1729 9408 ACGGTAGAGGCCCAAACGGC 1728 9409 CGTCCCGCGCAGGATCCAGTTGG 1800 9410 GTCCCGCGCAGGATCCAGTT 1801 9411 TCCCGCGCAGGATCCAGTTG 1802 9412 CCCGCGCAGGATCCAGTTGG 1803 9413 CCGCGCAGGATCCAGTTGGC 1804 9414 CGCGCAGGATCCAGTTGGCA 1805 9415 GCGCAGGATCCAGTTGGCAG 1806 9416 CGCAGGATCCAGTTGGCAGC 1807 9417 ACGTCCCGCGCAGGATCCAG 1799 9418 GACGTCCCGCGCAGGATCCA 1798 9419 GGACGTCCCGCGCAGGATCC 1797 9420 AGGACGTCCCGCGCAGGATC 1796 9421 AAGGACGTCCCGCGCAGGAT 1795 9422 AAAGGACGTCCCGCGCAGGA 1794 9423 CAAAGGACGTCCCGCGCAGG 1793 9424 ACAAAGGACGTCCCGCGCAG 1792 9425 GACAAAGGACGTCCCGCGCA 1791 9426 AGACAAAGGACGTCCCGCGC 1790 9427 TAGACAAAGGACGTCCCGCG 1789 9428 GTAGACAAAGGACGTCCCGC 1788 9429 CGTAGACAAAGGACGTCCCG 1787 9430 ACGTAGACAAAGGACGTCCC 1786 9431 GACGTAGACAAAGGACGTCC 1785 9432 CGGCTGCGAGCAAAACAAGCTGCTAG 1909 9433 GGCTGCGAGCAAAACAAGCT 1910 9434 GCTGCGAGCAAAACAAGCTG 1911 9435 CTGCGAGCAAAACAAGCTGC 1912 9436 TGCGAGCAAAACAAGCTGCT 1913 9437 GCGAGCAAAACAAGCTGCTA 1914 9438 CGAGCAAAACAAGCTGCTAG 1915 9439 CCGGCTGCGAGCAAAACAAG 1908 9440 ACCGGCTGCGAGCAAAACAA 1907 9441 GACCGGCTGCGAGCAAAACA 1906 9442 AGACCGGCTGCGAGCAAAAC 1905 9443 CAGACCGGCTGCGAGCAAAA 1904 9444 CCAGACCGGCTGCGAGCAAA 1903 9445 TCCAGACCGGCTGCGAGCAA 1902 9446 CTCCAGACCGGCTGCGAGCA 1901 9447 GCTCCAGACCGGCTGCGAGC 1900 9448 CGCTCCAGACCGGCTGCGAG 1899 9449 TCGCTCCAGACCGGCTGCGA 1898 9450 TTCGCTCCAGACCGGCTGCG 1897 9451 TTTCGCTCCAGACCGGCTGC 1896 9452 GTTTCGCTCCAGACCGGCTG 1895 9453 AGTTTCGCTCCAGACCGGCT 1894 9454 AAGTTTCGCTCCAGACCGGC 1893 9455 TAAGTTTCGCTCCAGACCGG 1892 9456 ATAAGTTTCGCTCCAGACCG 1891 9457 GATAAGTTTCGCTCCAGACC 1890 9458 CGATAAGTTTCGCTCCAGAC 1889 9459 CCGATAAGTTTCGCTCCAGA 1888 9460 TCCGATAAGTTTCGCTCCAG 1887 9461 TTCCGATAAGTTTCGCTCCA 1886 9462 GTTCCGATAAGTTTCGCTCC 1885 9463 GGTTCCGATAAGTTTCGCTC 1884 9464 CGGTTCCGATAAGTTTCGCT 1883 9465 TCGGTTCCGATAAGTTTCGC 1882 9466 GTCGGTTCCGATAAGTTTCG 1881 9467 TGTCGGTTCCGATAAGTTTC 1880 9468 CGCATGCGCCGATGGCCTATGGCCAA 1978 9469 GCATGCGCCGATGGCCTATG 1979 9470 CATGCGCCGATGGCCTATGG 1980 9471 ATGCGCCGATGGCCTATGGC 1981 9472 TGCGCCGATGGCCTATGGCC 1982 9473 GCGCCGATGGCCTATGGCCA 1983 9474 CGCCGATGGCCTATGGCCAA 1984 9475 GCCGATGGCCTATGGCCAAG 1985 9476 CCGATGGCCTATGGCCAAGC 1986 9477 CGATGGCCTATGGCCAAGCC 1987 9478 ACGCATGCGCCGATGGCCTA 1977 9479 CACGCATGCGCCGATGGCCT 1976 9480 CCACGCATGCGCCGATGGCC 1975 9481 TCCACGCATGCGCCGATGGC 1974 9482 TTCCACGCATGCGCCGATGG 1973 9483 GTTCCACGCATGCGCCGATG 1972 9484 GGTTCCACGCATGCGCCGAT 1971 9485 AGGTTCCACGCATGCGCCGA 1970 9486 AAGGTTCCACGCATGCGCCG 1969 9487 AAAGGTTCCACGCATGCGCC 1968 9488 CAAAGGTTCCACGCATGCGC 1967 9489 ACAAAGGTTCCACGCATGCG 1966 9490 CACAAAGGTTCCACGCATGC 1965 9491 CCACAAAGGTTCCACGCATG 1964 9492 GCCACAAAGGTTCCACGCAT 1963 9493 AGCCACAAAGGTTCCACGCA 1962 9494 GAGCCACAAAGGTTCCACGC 1961 9495 GGAGCCACAAAGGTTCCACG 1960 9496 CGCCGCAGACACATCCAGCGATA 2826 9497 GCCGCAGACACATCCAGCGA 2827 9498 CCGCAGACACATCCAGCGAT 2828 9499 CGCAGACACATCCAGCGATA 2829 9500 GCAGACACATCCAGCGATAG 2830 9501 CAGACACATCCAGCGATAGC 2831 9502 AGACACATCCAGCGATAGCC 2832 9503 GACACATCCAGCGATAGCCA 2833 9504 ACACATCCAGCGATAGCCAG 2834 9505 CACATCCAGCGATAGCCAGG 2835 9506 ACATCCAGCGATAGCCAGGA 2836 9507 CATCCAGCGATAGCCAGGAC 2837 9508 ATCCAGCGATAGCCAGGACA 2838 9509 TCCAGCGATAGCCAGGACAA 2839 9510 CCAGCGATAGCCAGGACAAG 2840 9511 CAGCGATAGCCAGGACAAGT 2841 9512 AGCGATAGCCAGGACAAGTT 2842 9513 GCGATAGCCAGGACAAGTTG 2843 9514 CGATAGCCAGGACAAGTTGG 2844 9515 ACGCCGCAGACACATCCAGC 2825 9516 AACGCCGCAGACACATCCAG 2824 9517 AAACGCCGCAGACACATCCA 2823 9518 AAAACGCCGCAGACACATCC 2822 9519 TAAAACGCCGCAGACACATC 2821 9520 ATAAAACGCCGCAGACACAT 2820 9521 GATAAAACGCCGCAGACACA 2819 9522 TGATAAAACGCCGCAGACAC 2818 9523 ATGATAAAACGCCGCAGACA 2817 9524 TATGATAAAACGCCGCAGAC 2816 9525 GCTCCAGACCGGCTGCGA 1900 9526 CTCCAGACCGGCTGCGAGCA 1901 9527 TCCAGACCGGCTGCGAGCAA 1902 9528 CCAGACCGGCTGCGAGCAAA 1903 9529 CAGACCGGCTGCGAGCAAAA 1904 9530 AGACCGGCTGCGAGCAAAAC 1905 9531 GACCGGCTGCGAGCAAAACA 1906 9532 ACCGGCTGCGAGCAAAACAA 1907 9533 CCGGCTGCGAGCAAAACAAG 1908 9534 CGGCTGCGAGCAAAACAAGC 1909 9535 GGCTGCGAGCAAAACAAGCT 1910 9536 GCTGCGAGCAAAACAAGCTG 1911 9537 CTGCGAGCAAAACAAGCTGC 1912 9538 TGCGAGCAAAACAAGCTGCT 1913 9539 GCGAGCAAAACAAGCTGCTA 1914 9540 CGAGCAAAACAAGCTGCTAG 1915 9541 CGCTCCAGACCGGCTGCGAG 1899 9542 TCGCTCCAGACCGGCTGCGA 1898 9543 TTCGCTCCAGACCGGCTGCG 1897 9544 TTTCGCTCCAGACCGGCTGC 1896 9545 GTTTCGCTCCAGACCGGCTG 1895 9546 AGTTTCGCTCCAGACCGGCT 1894 9547 AAGTTTCGCTCCAGACCGGC 1893 9548 TAAGTTTCGCTCCAGACCGG 1892 9549 ATAAGTTTCGCTCCAGACCG 1891 9550 GATAAGTTTCGCTCCAGACC 1890 9551 CGATAAGTTTCGCTCCAGAC 1889 9552 CCGATAAGTTTCGCTCCAGA 1888 9553 TCCGATAAGTTTCGCTCCAG 1887 9554 TTCCGATAAGTTTCGCTCCA 1886 9555 GTTCCGATAAGTTTCGCTCC 1885 9556 GGTTCCGATAAGTTTCGCTC 1884 9557 CGGTTCCGATAAGTTTCGCT 1883 9558 TCGGTTCCGATAAGTTTCGC 1882 9559 GTCGGTTCCGATAAGTTTCG 1881 9560 TGTCGGTTCCGATAAGTTTC 1880 9561 CGTCCATCGCAGGATCCAGTTGG 1800 9562 CGCCGCAGACACATCCAGCGATA 2826 9563 GCCGCAGACACATCCAGCGA 2827 9564 CCGCAGACACATCCAGCGAT 2828 9565 CGCAGACACATCCAGCGATA 2829 9566 GCAGACACATCCAGCGATAG 2830 9567 CAGACACATCCAGCGATAGC 2831 9568 AGACACATCCAGCGATAGCC 2832 9569 GACACATCCAGCGATAGCCA 2833 9570 ACACATCCAGCGATAGCCAG 2834 9571 CACATCCAGCGATAGCCAGG 2835 9572 ACATCCAGCGATAGCCAGGA 2836 9573 CATCCAGCGATAGCCAGGAC 2837 9574 ATCCAGCGATAGCCAGGACA 2838 9575 TCCAGCGATAGCCAGGACAA 2839 9576 CCAGCGATAGCCAGGACAAG 2840 9577 CAGCGATAGCCAGGACAAGT 2841 9578 AGCGATAGCCAGGACAAGTT 2842 9579 GCGATAGCCAGGACAAGTTG 2843 9580 CGATAGCCAGGACAAGTTGG 2844 9581 ACGCCGCAGACACATCCAGC 2825 9582 AACGCCGCAGACACATCCAG 2824 9583 AAACGCCGCAGACACATCCA 2823 9584 AAAACGCCGCAGACACATCC 2822 9585 TAAAACGCCGCAGACACATC 2821 9586 ATAAAACGCCGCAGACACAT 2820 9587 GATAAAACGCCGCAGACACA 2819 9588 TGATAAAACGCCGCAGACAC 2818 9589 ATGATAAAACGCCGCAGACA 2817 9590 TATGATAAAACGCCGCAGAC 2816 9591 CAAATGGCACTAGTAAACTGAG 2524 9592 GAGATTGAGATCTGCGGCGACGCGG 780 9593 CGACGCGGCGATTGAGATCTTCGTCTG 801 9594 AGGGGTCGTCCGCGGGATTCAGCGCCG 1754

Hot Zones (Relative upstream location to gene start site) 245-425 785-965 1145-1235 1505-2135 2585-3125

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11980) CTCCACAACATTCCACCAAGCTCTGCTAGATCCCAGAGTGAGGGGCCTAT ATTTTCCTGCTGGTGGCTCCAGTTCCGGAACAGTAAACCCTGTTCCGACT ACTGCCTCACCCATATCGTCAATCTTCTCGAGGACTGGGGACCCTGCACC GAACATGGAGAGCACAACATCAGGATTCCTAGGACCCCTGCTCGTGTTAC AGGCGGGGTTTTTCTTGTTGACAAGAATCCTCACAATACCACAGAGTCTA GACTCGTGGTGGACTTCTCTCAATTTTCTAGGGGGAGCACCCACGTGTCC TGGCCAAAATTCGCAGTCCCCAACCTCCAATCACTCACCAACCTCTTGTC CTCCAACTTGTCCTGGCTATCGCTGGATGTGTCTGCGGCGTTTTATCATA TTCCTCTTCATCCTGCTGCTATGCCTCATCTTCTTGTTGGTTCTTCTGGA CTACCAAGGTATGTTGCCCGTTTGTCCTCTACTTCCAGGAACATCAACTA CCAGCACGGGACCATGCAGAACCTGCACGATTCCTGCTCAAGGAACCTCT ATGTTTCCCTCTTGTTGCTGTACAAAACCTTCGGACGGAAACTGCACTTG TATTCCCATCCCATCATCCTGGGCTTTCGCAAGATTCCTATGGGAGTGGG CCTCAGTCCGTTTCTCCTGGCTCAGTTTACTAGTGCCATTTGTTCAGTGG TTCGTAGGGCTTTCCCCCACTGTTTGGCTTTCAGCTATATGGATGATGTG GTATTGGGGGCCAAGTCTGTACAACATCTTGAGTCCCTTTTTACCTCTAT TACCAATTTTCTTTTGTCTTTGGGTATACATTTGAACCCTAATAAAACCA AACGTTGGGGCTACTCCCTTAACTTCATGGGATATGTAATTGGAAGTTGG GGTACTTTACCGCAGGAACATATTGTACAAAAACTCAAGCAATGTTTTCG AAAATTGCCTGTAAATAGACCTATTGATTGGAAAGTATGTCAAAGAATTG TGGGTCTTTTGGGCTTTGCTGCCCCTTTTACACAATGTGGCTATCCTGCC TTGATGCCTTTATATGCATGTATACAATCTAAGCAGGCTTTCACTTTCTC GCCAACTTACAAGGCCTTTCTGTGTAAACAATATCTAAACCTTTACCCCG TTGCCCGGCAACGGTCAGGTCTCTGCCAAGTGTTTGCTGACGCAACCCCC ACGGGTTGGGGCTTGGCCATAGGCCATCGGCGCATGCGTGGAACCTTTGT GGCTCCTCTGCCGATCCATACTGCGGAACTCCTAGCAGCTTGTTTTGCTC GCAGCCGGTCTGGAGCGAAACTTATCGGAACCGACAACTCAGTTGTCCTC TCTCGGAAATACACCTCCTTTCCATGGCTGCTAGGCTGTGCTGCCAACTG GATCCTGCGCGGGACGTCCTTTGTCTACGTCCCGTCGGCGCTGAATCCCG CGGACGACCCGTCTCGGGGCCGTTTGGGCCTCTACCGTCCCCTTCTTCAT CTGCCGTTCCGGCCGACCACGGGGCGCACCTCTCTTTACGCGGTCTCCCC GTCTGTGCCTTCTCATCTGCCGGACCGTGTGCACTTCGCTTCACCTCTGC ACGTAGCATGGAGACCACCGTGAACGCCCACCAGGTCTTGCCCAAGGTCT TACACAAGAGGACTCTTGGACTCTCAGCAATGTCAACGACCGACCTTGAG GCATACTTCAAAGACTGTTTGTTTAAAGACTGGGAGGAGTTGGGGGAGGA GATTAGGTTAAAGGTCTTTGTACTAGGAGGCTGTAGGCATAAATTGGTCT GTTCACCAGCACCATGCAACTTTTTCCCCTCTGCCTAATCATCTCATGTT CATGTCCTACTGTTCAAGCCTCCAAGCTGTGCCTTGGGTGGCTTTGGGGC ATGGACATTGACCCGTATAAAGAATTTGGAGCTTCTGTGGAGTTACTCTC TTTTTTGCCTTCTGACTTCTTTCCTTCTATTCGAGATCTCCTCGACACCG CCTCTGCTCTGTATCGGGAGGCCTTAGAGTCTCCGGAACATTGTTCACCT CACCATACAGCACTCAGGCAAGCTATTCTGTGTTGGGGTGAGTTGATGAA TCTGGCCACCTGGGTGGGAAGTAATTTGGAAGACCCAGCATCCAGGGAAT TAGTAGTCAGCTATGTCAATGTTAATATGGGCCTAAAAATTAGACAACTA TTGTGGTTTCACATTTCCTGCCTTACTTTTGGAAGAGAAACTGTCCTTGA GTATTTGGTGTCTTTTGGAGTGTGGATTCGCACTCCTCCCGCTTACAGAC CACCAAATGCCCCTATCTTATCAACACTTCCGGAAACTACTGTTGTTAGA CGACGAGGCAGGTCCCCTAGAAGAAGAACTCCCTCGCCTCGCAGACGAAG GTCTCAATCGCCGCGTCGCAGAAGATCTCAATCTCGGGAATCTCAATGTT AGTATCCCTTGGACTCATAAGGTGGGAAACTTTACTGGGCTTTATTCTTC TACTGTACCTGTCTTTAATCCTGATTGGAAAACTCCCTCCTTTCCTCACA TTCATTTACAGGAGGACATTATTAATAGATGTCAACAATATGTGGGCCCT CTGACAGTTAATGAAAAAAGGAGATTAAAATTAATTATGCCTGCTAGGTT CTATCCTAACCTTACCAAATATTTGCCCTTGGACAAAGGCATTAAACCGT ATTATCCTGAATATGCAGTTAATCATTACTTCAAAACTAGGCATTATTTA CATACTCTGTGGAAGGCTGGCATTCTATATAAGAGAGAAACTACACGCAG CGCCTCATTTTGTGGGTCACCATATTCTTGGGAACAAGAGCTACAGCATG GGAGGTTGGTCTTCCAAACCTCGACAAGGCATGGGGACGAATCTTTCTGT TCCCAATCCTCTGGGATTCTTTCCCGATCACCAGTTGGACCCTGCGTTCG GAGCCAACTCAAACAATCCAGATTGGGACTTCAACCCCAACAAGGATCAC TGGCCAGAGGCAAATCAGGTAGGAGCGGGAGCATTTGGTCCAGGGTTCAC CCCACCACACGGAGGCCTTTTGGGGTGGAGCCCTCAGGCTCAGGGCATAT TGACAACACTGCCAGCAGCACCTCCTCCTGCCTCCACCAATCGGCAGTCA GGAAGACAGCCTACTCCCATCTCTCCACCTCTAAGAGACAGTCATCCTCA GGCCATGCAGTGGAA

PARP1

Poly [ADP-ribose] polymerase 1 (PARP-1) is an enzyme that in humans is encoded by the PARP1 gene. PARP1 works to on single strands of DNA, modifies nuclear proteins by poly ADP-ribosylation, involved in differentiation, proliferation and tumor transformation. PARP1 also has a role in repair of single-stranded DNA (ssDNA) breaks. Reducing intracellular PARP1 levels with siRNA or inhibiting PARP1 activity with small molecules reduces repair of ssDNA breaks. In the absence of PARP1, when these breaks are encountered during DNA replication, the replication fork stalls, and double-strand DNA (dsDNA) breaks accumulate. These dsDNA breaks are repaired via homologous recombination (HR) repair, a potentially error-free repair mechanism. However, both BRCA1 and BRCA2 are at least partially necessary for the HR pathway to function. Therefore, cells that are deficient in BRCA1 or BRCA2 have been shown to be highly sensitive to PARP1 inhibition or knock-down, resulting in cell death by apoptosis, in stark contrast to cells with at least one good copy of both BRCA1 and BRCA2. Many breast cancers have defects in the BRCA1/BRCA2 HR repair pathway due to mutations in either BRCA1 or BRCA2 (termed BRCAness), or other essential genes in the pathway and thus thought to be highly sensitive to PARP1 inhibitors. PARP1 inhibitors are believed to be effective for cancers with BRCAness, due to the high sensitivity of the tumors to the inhibitor and the lack of deleterious effects on the remaining healthy cells with functioning BRCA HR pathway (Bryant et al. (2005) Nature 434 (7035): 913-7 and Farmer et al. (2005) Nature 434 (7035): 917-21. This is in contrast to conventional chemotherapies, which are highly toxic to all cells and can induce DNA damage in healthy cells, leading to secondary cancer generation.

Protein: PARP1 Gene: PARP1: (Homo sapiens, chromosome 1, 226548392-226595801 [NCBI Reference Sequence NC_(—)000001.10]; start site location: 226595630; strand: negative)

Gene Identification GeneID 142 HGNC 270 MIM 173870

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 9595 CCGCCAAAGCTCCGGAAGCCCGACGCC 14 9741 CCGCCTCGCCGCCTCGCGTGCGCTC 60 9887 CGGGAACGCCCACGGAACCCGCGTC 177 9933 CGGGTGGAGCTCTGCGGGCCGCTGC 269 9992 CGCCGGCCCCAAACTCTTAAGTGTG 696 10014 CGGGAAGCGCAGGCCCCCGCCTCGG 749 10045 CGTTCTAACCTGCCGTCCACAGACC 839

Target Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 9595 CCGCCAAAGCTCCGGAAGCCCGACGCC 14 9596 CGCCAAAGCTCCGGAAGCCC 15 9597 GCCAAAGCTCCGGAAGCCCG 16 9598 CCAAAGCTCCGGAAGCCCGA 17 9599 CAAAGCTCCGGAAGCCCGAC 18 9600 AAAGCTCCGGAAGCCCGACG 19 9601 AAGCTCCGGAAGCCCGACGC 20 9602 AGCTCCGGAAGCCCGACGCC 21 9603 GCTCCGGAAGCCCGACGCCA 22 9604 CTCCGGAAGCCCGACGCCAC 23 9605 TCCGGAAGCCCGACGCCACG 24 9606 CCGGAAGCCCGACGCCACGA 25 9607 CGGAAGCCCGACGCCACGAC 26 9608 GGAAGCCCGACGCCACGACC 27 9609 GAAGCCCGACGCCACGACCT 28 9610 AAGCCCGACGCCACGACCTA 29 9611 AGCCCGACGCCACGACCTAG 30 9612 GCCCGACGCCACGACCTAGA 31 9613 CCCGACGCCACGACCTAGAA 32 9614 CCGACGCCACGACCTAGAAA 33 9615 CGACGCCACGACCTAGAAAC 34 9616 GACGCCACGACCTAGAAACA 35 9617 ACGCCACGACCTAGAAACAC 36 9618 CGCCACGACCTAGAAACACG 37 9619 GCCACGACCTAGAAACACGC 38 9620 CCACGACCTAGAAACACGCT 39 9621 CACGACCTAGAAACACGCTG 40 9622 ACGACCTAGAAACACGCTGC 41 9623 CGACCTAGAAACACGCTGCC 42 9624 GACCTAGAAACACGCTGCCG 43 9625 ACCTAGAAACACGCTGCCGC 44 9626 CCTAGAAACACGCTGCCGCC 45 9627 CTAGAAACACGCTGCCGCCT 46 9628 TAGAAACACGCTGCCGCCTC 47 9629 AGAAACACGCTGCCGCCTCG 48 9630 GAAACACGCTGCCGCCTCGC 49 9631 AAACACGCTGCCGCCTCGCC 50 9632 AACACGCTGCCGCCTCGCCG 51 9633 ACACGCTGCCGCCTCGCCGC 52 9634 CACGCTGCCGCCTCGCCGCC 53 9635 ACGCTGCCGCCTCGCCGCCT 54 9636 CGCTGCCGCCTCGCCGCCTC 55 9637 GCTGCCGCCTCGCCGCCTCG 56 9638 CTGCCGCCTCGCCGCCTCGC 57 9639 TGCCGCCTCGCCGCCTCGCG 58 9640 GCCGCCTCGCCGCCTCGCGT 59 9641 CCGCCTCGCCGCCTCGCGTG 60 9642 CGCCTCGCCGCCTCGCGTGC 61 9643 GCCTCGCCGCCTCGCGTGCG 62 9644 CCTCGCCGCCTCGCGTGCGC 63 9645 CTCGCCGCCTCGCGTGCGCT 64 9646 TCGCCGCCTCGCGTGCGCTC 65 9647 CGCCGCCTCGCGTGCGCTCA 66 9648 GCCGCCTCGCGTGCGCTCAC 67 9649 CCGCCTCGCGTGCGCTCACC 68 9650 CGCCTCGCGTGCGCTCACCC 69 9651 GCCTCGCGTGCGCTCACCCA 70 9652 CCTCGCGTGCGCTCACCCAG 71 9653 CTCGCGTGCGCTCACCCAGC 72 9654 TCGCGTGCGCTCACCCAGCC 73 9655 CGCGTGCGCTCACCCAGCCG 74 9656 GCGTGCGCTCACCCAGCCGC 75 9657 CGTGCGCTCACCCAGCCGCA 76 9658 GTGCGCTCACCCAGCCGCAG 77 9659 TGCGCTCACCCAGCCGCAGG 78 9660 GCGCTCACCCAGCCGCAGGC 79 9661 CGCTCACCCAGCCGCAGGCG 80 9662 GCTCACCCAGCCGCAGGCGC 81 9663 CTCACCCAGCCGCAGGCGCC 82 9664 TCACCCAGCCGCAGGCGCCT 83 9665 CACCCAGCCGCAGGCGCCTG 84 9666 ACCCAGCCGCAGGCGCCTGA 85 9667 CCCAGCCGCAGGCGCCTGAG 86 9668 CCAGCCGCAGGCGCCTGAGC 87 9669 CAGCCGCAGGCGCCTGAGCG 88 9670 AGCCGCAGGCGCCTGAGCGG 89 9671 GCCGCAGGCGCCTGAGCGGC 90 9672 CCGCAGGCGCCTGAGCGGCC 91 9673 CGCAGGCGCCTGAGCGGCCA 92 9674 GCAGGCGCCTGAGCGGCCAG 93 9675 CAGGCGCCTGAGCGGCCAGA 94 9676 AGGCGCCTGAGCGGCCAGAG 95 9677 GGCGCCTGAGCGGCCAGAGC 96 9678 GCGCCTGAGCGGCCAGAGCC 97 9679 CGCCTGAGCGGCCAGAGCCG 98 9680 GCCTGAGCGGCCAGAGCCGC 99 9681 CCTGAGCGGCCAGAGCCGCC 100 9682 CTGAGCGGCCAGAGCCGCCA 101 9683 TGAGCGGCCAGAGCCGCCAC 102 9684 GAGCGGCCAGAGCCGCCACC 103 9685 AGCGGCCAGAGCCGCCACCG 104 9686 GCGGCCAGAGCCGCCACCGA 105 9687 CGGCCAGAGCCGCCACCGAA 106 9688 GGCCAGAGCCGCCACCGAAC 107 9689 GCCAGAGCCGCCACCGAACA 108 9690 CCAGAGCCGCCACCGAACAC 109 9691 CAGAGCCGCCACCGAACACG 110 9692 AGAGCCGCCACCGAACACGC 111 9693 GAGCCGCCACCGAACACGCC 112 9694 AGCCGCCACCGAACACGCCG 113 9695 GCCGCCACCGAACACGCCGC 114 9696 CCGCCACCGAACACGCCGCA 115 9697 CGCCACCGAACACGCCGCAC 116 9698 GCCACCGAACACGCCGCACC 117 9699 CCACCGAACACGCCGCACCG 118 9700 CACCGAACACGCCGCACCGG 119 9701 ACCGAACACGCCGCACCGGC 120 9702 CCGAACACGCCGCACCGGCC 121 9703 CGAACACGCCGCACCGGCCA 122 9704 GAACACGCCGCACCGGCCAC 123 9705 AACACGCCGCACCGGCCACC 124 9706 ACACGCCGCACCGGCCACCG 125 9707 CACGCCGCACCGGCCACCGC 126 9708 ACGCCGCACCGGCCACCGCC 127 9709 CGCCGCACCGGCCACCGCCG 128 9710 GCCGCACCGGCCACCGCCGT 129 9711 CCGCACCGGCCACCGCCGTT 130 9712 CGCACCGGCCACCGCCGTTC 131 9713 GCACCGGCCACCGCCGTTCC 132 9714 CACCGGCCACCGCCGTTCCC 133 9715 ACCGGCCACCGCCGTTCCCT 134 9716 CCGGCCACCGCCGTTCCCTG 135 9717 CGGCCACCGCCGTTCCCTGA 136 9718 GGCCACCGCCGTTCCCTGAT 137 9719 GCCACCGCCGTTCCCTGATA 138 9720 CCACCGCCGTTCCCTGATAG 139 9721 CACCGCCGTTCCCTGATAGA 140 9722 ACCGCCGTTCCCTGATAGAT 141 9723 CCGCCGTTCCCTGATAGATT 142 9724 CGCCGTTCCCTGATAGATTG 143 9725 GCCGTTCCCTGATAGATTGC 144 9726 CCGTTCCCTGATAGATTGCT 145 9727 CGTTCCCTGATAGATTGCTG 146 9728 GCCGCCAAAGCTCCGGAAGC 13 9729 TGCCGCCAAAGCTCCGGAAG 12 9730 CTGCCGCCAAAGCTCCGGAA 11 9731 GCTGCCGCCAAAGCTCCGGA 10 9732 AGCTGCCGCCAAAGCTCCGG 9 9733 TAGCTGCCGCCAAAGCTCCG 8 9734 CTAGCTGCCGCCAAAGCTCC 7 9735 CCTAGCTGCCGCCAAAGCTC 6 9736 CCCTAGCTGCCGCCAAAGCT 5 9737 CCCCTAGCTGCCGCCAAAGC 4 9738 TCCCCTAGCTGCCGCCAAAG 3 9739 CTCCCCTAGCTGCCGCCAAA 2 9740 CCTCCCCTAGCTGCCGCCAA 1 9741 CCGCCTCGCCGCCTCGCGTGCGCTC 60 9742 CGCCTCGCCGCCTCGCGTGC 61 9743 GCCTCGCCGCCTCGCGTGCG 62 9744 CCTCGCCGCCTCGCGTGCGC 63 9745 CTCGCCGCCTCGCGTGCGCT 64 9746 TCGCCGCCTCGCGTGCGCTC 65 9747 CGCCGCCTCGCGTGCGCTCA 66 9748 GCCGCCTCGCGTGCGCTCAC 67 9749 CCGCCTCGCGTGCGCTCACC 68 9750 CGCCTCGCGTGCGCTCACCC 69 9751 GCCTCGCGTGCGCTCACCCA 70 9752 CCTCGCGTGCGCTCACCCAG 71 9753 CTCGCGTGCGCTCACCCAGC 72 9754 TCGCGTGCGCTCACCCAGCC 73 9755 CGCGTGCGCTCACCCAGCCG 74 9756 GCGTGCGCTCACCCAGCCGC 75 9757 CGTGCGCTCACCCAGCCGCA 76 9758 GTGCGCTCACCCAGCCGCAG 77 9759 TGCGCTCACCCAGCCGCAGG 78 9760 GCGCTCACCCAGCCGCAGGC 79 9761 CGCTCACCCAGCCGCAGGCG 80 9762 GCTCACCCAGCCGCAGGCGC 81 9763 CTCACCCAGCCGCAGGCGCC 82 9764 TCACCCAGCCGCAGGCGCCT 83 9765 CACCCAGCCGCAGGCGCCTG 84 9766 ACCCAGCCGCAGGCGCCTGA 85 9767 CCCAGCCGCAGGCGCCTGAG 86 9768 CCAGCCGCAGGCGCCTGAGC 87 9769 CAGCCGCAGGCGCCTGAGCG 88 9770 AGCCGCAGGCGCCTGAGCGG 89 9771 GCCGCAGGCGCCTGAGCGGC 90 9772 CCGCAGGCGCCTGAGCGGCC 91 9773 CGCAGGCGCCTGAGCGGCCA 92 9774 GCAGGCGCCTGAGCGGCCAG 93 9775 CAGGCGCCTGAGCGGCCAGA 94 9776 AGGCGCCTGAGCGGCCAGAG 95 9777 GGCGCCTGAGCGGCCAGAGC 96 9778 GCGCCTGAGCGGCCAGAGCC 97 9779 CGCCTGAGCGGCCAGAGCCG 98 9780 GCCTGAGCGGCCAGAGCCGC 99 9781 CCTGAGCGGCCAGAGCCGCC 100 9782 CTGAGCGGCCAGAGCCGCCA 101 9783 TGAGCGGCCAGAGCCGCCAC 102 9784 GAGCGGCCAGAGCCGCCACC 103 9785 AGCGGCCAGAGCCGCCACCG 104 9786 GCGGCCAGAGCCGCCACCGA 105 9787 CGGCCAGAGCCGCCACCGAA 106 9788 GGCCAGAGCCGCCACCGAAC 107 9789 GCCAGAGCCGCCACCGAACA 108 9790 CCAGAGCCGCCACCGAACAC 109 9791 CAGAGCCGCCACCGAACACG 110 9792 AGAGCCGCCACCGAACACGC 111 9793 GAGCCGCCACCGAACACGCC 112 9794 AGCCGCCACCGAACACGCCG 113 9795 GCCGCCACCGAACACGCCGC 114 9796 CCGCCACCGAACACGCCGCA 115 9797 CGCCACCGAACACGCCGCAC 116 9798 GCCACCGAACACGCCGCACC 117 9799 CCACCGAACACGCCGCACCG 118 9800 CACCGAACACGCCGCACCGG 119 9801 ACCGAACACGCCGCACCGGC 120 9802 CCGAACACGCCGCACCGGCC 121 9803 CGAACACGCCGCACCGGCCA 122 9804 GAACACGCCGCACCGGCCAC 123 9805 AACACGCCGCACCGGCCACC 124 9806 ACACGCCGCACCGGCCACCG 125 9807 CACGCCGCACCGGCCACCGC 126 9808 ACGCCGCACCGGCCACCGCC 127 9809 CGCCGCACCGGCCACCGCCG 128 9810 GCCGCACCGGCCACCGCCGT 129 9811 CCGCACCGGCCACCGCCGTT 130 9812 CGCACCGGCCACCGCCGTTC 131 9813 GCACCGGCCACCGCCGTTCC 132 9814 CACCGGCCACCGCCGTTCCC 133 9815 ACCGGCCACCGCCGTTCCCT 134 9816 CCGGCCACCGCCGTTCCCTG 135 9817 CGGCCACCGCCGTTCCCTGA 136 9818 GGCCACCGCCGTTCCCTGAT 137 9819 GCCACCGCCGTTCCCTGATA 138 9820 CCACCGCCGTTCCCTGATAG 139 9821 CACCGCCGTTCCCTGATAGA 140 9822 ACCGCCGTTCCCTGATAGAT 141 9823 CCGCCGTTCCCTGATAGATT 142 9824 CGCCGTTCCCTGATAGATTG 143 9825 GCCGTTCCCTGATAGATTGC 144 9826 CCGTTCCCTGATAGATTGCT 145 9827 CGTTCCCTGATAGATTGCTG 146 9828 GCCGCCTCGCCGCCTCGCGT 59 9829 TGCCGCCTCGCCGCCTCGCG 58 9830 CTGCCGCCTCGCCGCCTCGC 57 9831 GCTGCCGCCTCGCCGCCTCG 56 9832 CGCTGCCGCCTCGCCGCCTC 55 9833 ACGCTGCCGCCTCGCCGCCT 54 9834 CACGCTGCCGCCTCGCCGCC 53 9835 ACACGCTGCCGCCTCGCCGC 52 9836 AACACGCTGCCGCCTCGCCG 51 9837 AAACACGCTGCCGCCTCGCC 50 9838 GAAACACGCTGCCGCCTCGC 49 9839 AGAAACACGCTGCCGCCTCG 48 9840 TAGAAACACGCTGCCGCCTC 47 9841 CTAGAAACACGCTGCCGCCT 46 9842 CCTAGAAACACGCTGCCGCC 45 9843 ACCTAGAAACACGCTGCCGC 44 9844 GACCTAGAAACACGCTGCCG 43 9845 CGACCTAGAAACACGCTGCC 42 9846 ACGACCTAGAAACACGCTGC 41 9847 CACGACCTAGAAACACGCTG 40 9848 CCACGACCTAGAAACACGCT 39 9849 GCCACGACCTAGAAACACGC 38 9850 CGCCACGACCTAGAAACACG 37 9851 ACGCCACGACCTAGAAACAC 36 9852 GACGCCACGACCTAGAAACA 35 9853 CGACGCCACGACCTAGAAAC 34 9854 CCGACGCCACGACCTAGAAA 33 9855 CCCGACGCCACGACCTAGAA 32 9856 GCCCGACGCCACGACCTAGA 31 9857 AGCCCGACGCCACGACCTAG 30 9858 AAGCCCGACGCCACGACCTA 29 9859 GAAGCCCGACGCCACGACCT 28 9860 GGAAGCCCGACGCCACGACC 27 9861 CGGAAGCCCGACGCCACGAC 26 9862 CCGGAAGCCCGACGCCACGA 25 9863 TCCGGAAGCCCGACGCCACG 24 9864 CTCCGGAAGCCCGACGCCAC 23 9865 GCTCCGGAAGCCCGACGCCA 22 9866 AGCTCCGGAAGCCCGACGCC 21 9867 AAGCTCCGGAAGCCCGACGC 20 9868 AAAGCTCCGGAAGCCCGACG 19 9869 CAAAGCTCCGGAAGCCCGAC 18 9870 CCAAAGCTCCGGAAGCCCGA 17 9871 GCCAAAGCTCCGGAAGCCCG 16 9872 CGCCAAAGCTCCGGAAGCCC 15 9873 CCGCCAAAGCTCCGGAAGCC 14 9874 GCCGCCAAAGCTCCGGAAGC 13 9875 TGCCGCCAAAGCTCCGGAAG 12 9876 CTGCCGCCAAAGCTCCGGAA 11 9877 GCTGCCGCCAAAGCTCCGGA 10 9878 AGCTGCCGCCAAAGCTCCGG 9 9879 TAGCTGCCGCCAAAGCTCCG 8 9880 CTAGCTGCCGCCAAAGCTCC 7 9881 CCTAGCTGCCGCCAAAGCTC 6 9882 CCCTAGCTGCCGCCAAAGCT 5 9883 CCCCTAGCTGCCGCCAAAGC 4 9884 TCCCCTAGCTGCCGCCAAAG 3 9885 CTCCCCTAGCTGCCGCCAAA 2 9886 CCTCCCCTAGCTGCCGCCAA 1 9887 CGGGAACGCCCACGGAACCCGCGTC 177 9888 GGGAACGCCCACGGAACCCG 178 9889 GGAACGCCCACGGAACCCGC 179 9890 GAACGCCCACGGAACCCGCG 180 9891 AACGCCCACGGAACCCGCGT 181 9892 ACGCCCACGGAACCCGCGTC 182 9893 CGCCCACGGAACCCGCGTCC 183 9894 GCCCACGGAACCCGCGTCCA 184 9895 CCCACGGAACCCGCGTCCAC 185 9896 CCACGGAACCCGCGTCCACG 186 9897 CACGGAACCCGCGTCCACGG 187 9898 ACGGAACCCGCGTCCACGGG 188 9899 CGGAACCCGCGTCCACGGGG 189 9900 GGAACCCGCGTCCACGGGGC 190 9901 GAACCCGCGTCCACGGGGCG 191 9902 AACCCGCGTCCACGGGGCGG 192 9903 ACCCGCGTCCACGGGGCGGG 193 9904 CCCGCGTCCACGGGGCGGGG 194 9905 CCGCGTCCACGGGGCGGGGC 195 9906 CGCGTCCACGGGGCGGGGCC 196 9907 GCGTCCACGGGGCGGGGCCG 197 9908 CGTCCACGGGGCGGGGCCGG 198 9909 GTCCACGGGGCGGGGCCGGC 199 9910 TCCACGGGGCGGGGCCGGCG 200 9911 CCACGGGGCGGGGCCGGCGG 201 9912 CACGGGGCGGGGCCGGCGGC 202 9913 GCGGGAACGCCCACGGAACC 176 9914 CGCGGGAACGCCCACGGAAC 175 9915 CCGCGGGAACGCCCACGGAA 174 9916 GCCGCGGGAACGCCCACGGA 173 9917 GGCCGCGGGAACGCCCACGG 172 9918 TGGCCGCGGGAACGCCCACG 171 9919 CTGGCCGCGGGAACGCCCAC 170 9920 CCTGGCCGCGGGAACGCCCA 169 9921 GCCTGGCCGCGGGAACGCCC 168 9922 TGCCTGGCCGCGGGAACGCC 167 9923 ATGCCTGGCCGCGGGAACGC 166 9924 GATGCCTGGCCGCGGGAACG 165 9925 TGATGCCTGGCCGCGGGAAC 164 9926 CTGATGCCTGGCCGCGGGAA 163 9927 GCTGATGCCTGGCCGCGGGA 162 9928 TGCTGATGCCTGGCCGCGGG 161 9929 TTGCTGATGCCTGGCCGCGG 160 9930 ATTGCTGATGCCTGGCCGCG 159 9931 GATTGCTGATGCCTGGCCGC 158 9932 AGATTGCTGATGCCTGGCCG 157 9933 CGGGTGGAGCTCTGCGGGCCGCTGC 269 9934 GGGTGGAGCTCTGCGGGCCG 270 9935 GGTGGAGCTCTGCGGGCCGC 271 9936 GTGGAGCTCTGCGGGCCGCT 272 9937 TGGAGCTCTGCGGGCCGCTG 273 9938 GGAGCTCTGCGGGCCGCTGC 274 9939 GAGCTCTGCGGGCCGCTGCC 275 9940 AGCTCTGCGGGCCGCTGCCC 276 9941 GCTCTGCGGGCCGCTGCCCT 277 9942 CTCTGCGGGCCGCTGCCCTG 278 9943 TCTGCGGGCCGCTGCCCTGG 279 9944 CTGCGGGCCGCTGCCCTGGG 280 9945 TGCGGGCCGCTGCCCTGGGG 281 9946 GCGGGCCGCTGCCCTGGGGG 282 9947 CGGGCCGCTGCCCTGGGGGC 283 9948 GGGCCGCTGCCCTGGGGGCC 284 9949 GGCCGCTGCCCTGGGGGCCG 285 9950 GCCGCTGCCCTGGGGGCCGA 286 9951 CCGCTGCCCTGGGGGCCGAG 287 9952 CGCTGCCCTGGGGGCCGAGG 288 9953 GCTGCCCTGGGGGCCGAGGC 289 9954 CTGCCCTGGGGGCCGAGGCG 290 9955 TGCCCTGGGGGCCGAGGCGG 291 9956 GCCCTGGGGGCCGAGGCGGG 292 9957 CCCTGGGGGCCGAGGCGGGG 293 9958 CCTGGGGGCCGAGGCGGGGC 294 9959 CTGGGGGCCGAGGCGGGGCT 295 9960 TGGGGGCCGAGGCGGGGCTT 296 9961 CCGGGTGGAGCTCTGCGGGC 268 9962 GCCGGGTGGAGCTCTGCGGG 267 9963 TGCCGGGTGGAGCTCTGCGG 266 9964 CTGCCGGGTGGAGCTCTGCG 265 9965 CCTGCCGGGTGGAGCTCTGC 264 9966 GCCTGCCGGGTGGAGCTCTG 263 9967 CGCCTGCCGGGTGGAGCTCT 262 9968 GCGCCTGCCGGGTGGAGCTC 261 9969 GGCGCCTGCCGGGTGGAGCT 260 9970 GGGCGCCTGCCGGGTGGAGC 259 9971 CGGGCGCCTGCCGGGTGGAG 258 9972 CCGGGCGCCTGCCGGGTGGA 257 9973 CCCGGGCGCCTGCCGGGTGG 256 9974 TCCCGGGCGCCTGCCGGGTG 255 9975 TTCCCGGGCGCCTGCCGGGT 254 9976 TTTCCCGGGCGCCTGCCGGG 253 9977 GTTTCCCGGGCGCCTGCCGG 252 9978 AGTTTCCCGGGCGCCTGCCG 251 9979 GAGTTTCCCGGGCGCCTGCC 250 9980 GGAGTTTCCCGGGCGCCTGC 249 9981 CGGAGTTTCCCGGGCGCCTG 248 9982 GCGGAGTTTCCCGGGCGCCT 247 9983 GGCGGAGTTTCCCGGGCGCC 246 9984 GGGCGGAGTTTCCCGGGCGC 245 9985 GGGGCGGAGTTTCCCGGGCG 244 9986 GGGGGCGGAGTTTCCCGGGC 243 9987 GGGGGGCGGAGTTTCCCGGG 242 9988 CGGGGGGCGGAGTTTCCCGG 241 9989 CCGGGGGGCGGAGTTTCCCG 240 9990 GCCGGGGGGCGGAGTTTCCC 239 9991 GGCCGGGGGGCGGAGTTTCC 238 9992 CGCCGGCCCCAAACTCTTAAGTGTG 696 9993 GCCGGCCCCAAACTCTTAAG 697 9994 CCGGCCCCAAACTCTTAAGT 698 9995 CGGCCCCAAACTCTTAAGTG 699 9996 ACGCCGGCCCCAAACTCTTA 695 9997 CACGCCGGCCCCAAACTCTT 694 9998 CCACGCCGGCCCCAAACTCT 693 9999 ACCACGCCGGCCCCAAACTC 692 10000 TACCACGCCGGCCCCAAACT 691 10001 CTACCACGCCGGCCCCAAAC 690 10002 GCTACCACGCCGGCCCCAAA 689 10003 AGCTACCACGCCGGCCCCAA 688 10004 GAGCTACCACGCCGGCCCCA 687 10005 TGAGCTACCACGCCGGCCCC 686 10006 ATGAGCTACCACGCCGGCCC 685 10007 CATGAGCTACCACGCCGGCC 684 10008 GCATGAGCTACCACGCCGGC 683 10009 GGCATGAGCTACCACGCCGG 682 10010 GGGCATGAGCTACCACGCCG 681 10011 GGGGCATGAGCTACCACGCC 680 10012 AGGGGCATGAGCTACCACGC 679 10013 CAGGGGCATGAGCTACCACG 678 10014 CGGGAAGCGCAGGCCCCCGCCTCGG 749 10015 GGGAAGCGCAGGCCCCCGCC 750 10016 GGAAGCGCAGGCCCCCGCCT 751 10017 GAAGCGCAGGCCCCCGCCTC 752 10018 AAGCGCAGGCCCCCGCCTCG 753 10019 AGCGCAGGCCCCCGCCTCGG 754 10020 GCGCAGGCCCCCGCCTCGGG 755 10021 CGCAGGCCCCCGCCTCGGGA 756 10022 GCAGGCCCCCGCCTCGGGAA 757 10023 CAGGCCCCCGCCTCGGGAAT 758 10024 AGGCCCCCGCCTCGGGAATA 759 10025 GGCCCCCGCCTCGGGAATAT 760 10026 GCCCCCGCCTCGGGAATATA 761 10027 CCCCCGCCTCGGGAATATAG 762 10028 CCCCGCCTCGGGAATATAGT 763 10029 CCCGCCTCGGGAATATAGTT 764 10030 CCGCCTCGGGAATATAGTTG 765 10031 CGCCTCGGGAATATAGTTGA 766 10032 GCCTCGGGAATATAGTTGAT 767 10033 CCGGGAAGCGCAGGCCCCCG 748 10034 TCCGGGAAGCGCAGGCCCCC 747 10035 GTCCGGGAAGCGCAGGCCCC 746 10036 GGTCCGGGAAGCGCAGGCCC 745 10037 GGGTCCGGGAAGCGCAGGCC 744 10038 TGGGTCCGGGAAGCGCAGGC 743 10039 CTGGGTCCGGGAAGCGCAGG 742 10040 GCTGGGTCCGGGAAGCGCAG 741 10041 AGCTGGGTCCGGGAAGCGCA 740 10042 CAGCTGGGTCCGGGAAGCGC 739 10043 GCAGCTGGGTCCGGGAAGCG 738 10044 GGCAGCTGGGTCCGGGAAGC 737 10045 CGTTCTAACCTGCCGTCCACAGACC 839 10046 GTTCTAACCTGCCGTCCACA 840 10047 TTCTAACCTGCCGTCCACAG 841 10048 TCTAACCTGCCGTCCACAGA 842 10049 CTAACCTGCCGTCCACAGAC 843 10050 TAACCTGCCGTCCACAGACC 844 10051 AACCTGCCGTCCACAGACCG 845 10052 ACCTGCCGTCCACAGACCGT 846 10053 CCTGCCGTCCACAGACCGTC 847 10054 CTGCCGTCCACAGACCGTCG 848 10055 TGCCGTCCACAGACCGTCGG 849 10056 GCCGTCCACAGACCGTCGGG 850 10057 CCGTCCACAGACCGTCGGGA 851 10058 CGTCCACAGACCGTCGGGAC 852 10059 GTCCACAGACCGTCGGGACA 853 10060 TCCACAGACCGTCGGGACAA 854 10061 CCACAGACCGTCGGGACAAA 855 10062 CACAGACCGTCGGGACAAAA 856 10063 ACAGACCGTCGGGACAAAAT 857 10064 CAGACCGTCGGGACAAAATA 858 10065 AGACCGTCGGGACAAAATAC 859 10066 GACCGTCGGGACAAAATACC 860 10067 ACCGTCGGGACAAAATACCA 861 10068 CCGTCGGGACAAAATACCAA 862 10069 CGTCGGGACAAAATACCAAC 863 10070 GTCGGGACAAAATACCAACT 864 10071 TCGGGACAAAATACCAACTG 865 10072 CGGGACAAAATACCAACTGA 866 10073 GCGTTCTAACCTGCCGTCCA 838 10074 GGCGTTCTAACCTGCCGTCC 837 10075 GGGCGTTCTAACCTGCCGTC 836 10076 CGGGCGTTCTAACCTGCCGT 835 10077 ACGGGCGTTCTAACCTGCCG 834 10078 GACGGGCGTTCTAACCTGCC 833 10079 GGACGGGCGTTCTAACCTGC 832 10080 TGGACGGGCGTTCTAACCTG 831 10081 TTGGACGGGCGTTCTAACCT 830 10082 CTTGGACGGGCGTTCTAACC 829 10083 GCTTGGACGGGCGTTCTAAC 828 10084 GGCTTGGACGGGCGTTCTAA 827 10085 TGGCTTGGACGGGCGTTCTA 826 10086 CTGGCTTGGACGGGCGTTCT 825 10087 CCTGGCTTGGACGGGCGTTC 824 10088 TCCTGGCTTGGACGGGCGTT 823 10089 CTCCTGGCTTGGACGGGCGT 822 10090 CCTCCTGGCTTGGACGGGCG 821 10091 CCCTCCTGGCTTGGACGGGC 820 10092 ACCCTCCTGGCTTGGACGGG 819 10093 CACCCTCCTGGCTTGGACGG 818 10094 CCACCCTCCTGGCTTGGACG 817

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11981) GGTGGATCTCCACATGCAGAAGAATGTAGCTGGACCCATACCTTACACCA AATGTTTGTTGTGAGTTTATTTACTTTTTTGTGTGTGTGGAGACAGGGTC GTGCTATGTTGTCCAGGCTGATCTAGAACTCCTTACCTAGAGACACTGCC AAGGTAAGTGAGGGCCAAGTGGACACTGAGTGATTCTGTGCCTCACTGAG CAAAAATAACTAAACATGGGCGAAGGAGAGCCCAATGATCCCAGGGACAA AATGTCATCACGGGCATTCTGCGCACGCTTGCCAGGATACAGGAGAAGCA ACCAGACACTTCATTCATCTTCTCAGAATGTTCATTAACATGTTCAGAAA GGTGGAAAACCTTACTTGCTAAAGAGAAGGAAATTGGAGGCATGGCCAAA AGTATTCAAGGCCCTTTATGAAAAAGAAATGAAAACTGATATCCCTCCTA AAAGAGAAGTAAAACAGAAATTCAGAGATTCTAATGCACCCGAAAGGCCT CCTTTGGGCTTTCACTTTGTGTTCTGAGTACTGCCCTCAAATCAAAGGAG ATCCCGGTCTGTCCACTGGCAGTGATGCCAAGAACCTGGGAGGGACATGA GTGACCATGCTGCAGATGGCAAGCAGCCCAAAAAGAAGGCTTCTCAACTG AAGGAAAAGTACCAAGAGCAGAATGCTGCATATCCAGCCAAAGGAAAGCT GATGTGGCAAAAATGATGCTGTCAAGGCCGAAAAAGGCAAGAAAAAAAAA CAAAGCGGAGAAAGACAGAGAAGGTAAGGAAAATAAAAAATGAAGTCGAT GATAATGACAAATAAGGTGGTTCTATGGCAGCTTTTTTTTTTTTCTCTTG TCTATAAAGCATTTAACCTACCTGGACACAGCTCATTCCTTTTAAAGAAA AAAATTGAAATGTAAAGCCACCTAAGATTTATTTGTAAACTGCATGATGG CGTTCTTTTTCTGTTTTTGTATTATTAACAAGAATTATCAAGTAATTCTT CAGACAACCCTGTCCTGGTGGTATTTTGTATAGCCACCAACTTTGCCTGG TATACTATAGGGGTTATAAATCAGCATGGGAATTTCAAATTTAAGGCACA GTATAAGTTAGTTATATACAAATGTGAAGTAACATTATTAATTAAACTGT TGGCCTGTGCGAAGGGAGGGCCAACTGTGGGATTCAGTCATTCATTCAAC AAATATTGGTGAGTGCCTGACACTGTTCCAGGCACTGAGGCTATTGCAAC AAAACAGACACAAGCTCCTGCCCTCATGGAGCTTACATTCTGGTGAGGGA TACAGAGCCACCAAAAAGGATGGCAGCTGGGCCATGAGAAAGGATCAAAG TCAGGAAGTTAGAATTCGGGGATGGATTGAACATGGGACAAAAGAGAAGA GTCAAGTTGACTACAAAGCATTTGGCCTAAGTAATGCAAAGAATGGTGGG CCATTTCCTGAGATGGGAAGCACTAGGGTAGTTTTGGACATAAATGGAGA TGCATATAAGCCATCCAAACTGAAATATTGAGAAGGCAGTAGGTGATAGT TGGCTTTCCTTAGGTTCTAGGGCAGGAATTCTTAACCTTTTGTGTGTGTG CCTAGGACCCCTTTGGTGGTCCATGAAGCCCTTTCCAGAATAAATATTGT GGAGGAACCTACCTTAATGCAATAGTAGCTTCTAGGTACATTATCAGGCA AACTATCCCACAAGTTACAAAACAGAAAGCCTCACAGACCAAATTATGAT GCTTGAATTGCAGGGTTTATTGAATCAGTTTAAAACCACTTACAGCAAGA ACTCGATGGGGTGCATAACATACACAGGATAGGGTACAGGCGAGGCAGAT GGACCACACCACCAGAACCTAGAATTAGGGAATCCTCCCCTCCCCTCCCC TCCCACCCCTCCCCCCTTCCCCCTCCCCTCCCCTCCCCTCCTCCCCTCCT CTCCCATCCTCCCCTCCCCTCCCATCCTCCCCTCCCCTCCCCTTTTCTCT TCTTTTCTTTTTTTGAGACTGTCTCACTATGTTGCCCAGGCTGGAGTGCA ATGGCGTGATCTCGGCTCACTGCAACTTCCACCTCCTGGGTTCAAGCGAT TCTCATGCCTCAGCCTCCCGAGTAGCTGGGATTACAGGCACGCACCACAA TGCCCAGCTAATTTTTGTATTCTTAGTAGAGACGGGGTTTCACCATGTTG ACCAGGCTGGTTTTGAACTCCTGACCTTAGGTGATCCACCCGCCTCAGAT TCCCAAAGCGCTGGGATTACAGGCATGAGCCACTGCACCTGGCTAATATT GATATGTTTTCCCTCTCTCTGCCGCATCAGCCTGTCCCACTGACAGAGTT GAGGATGCTCAAGGCGGCTCAACAGAGGGTACCTGGAGCAACTCACACTG CACTATCAGAGAGACACAAGTGCAAGCACACTCAGCCACAGCTGCAGCTC ACCAATCAGCCTGCTGAACAGACCTGAACTTTAGCTGCATTTTTGGGGCA GAGCATATGGGTGCCAGGATGGGACCATAATCTTATCACCAATGAGTGGC CATTTAGGGATGATATAGTTGTCAACCCAGAGATGGCATGATCATGCCTT TTGACTTGGTCATTCTCTAAGTAAAACTTTTATTTGTTCCATCATATTTT CCACTTATTCTGTTTACCTTCAAAATATCTTTTTTTTTTTTTTTTGAGAC AGGGTCACACTGTCACCCAGGCTAGAGTCCAGTGGCACTATCATGGCTCA CCACAGCCTCAACCTTCAGGGCTCAGGTGATCCTCCCACTTCAGCCTCCC GAGTAGATGGGACTACAGGCACCTGCCACCACCCCCAGCTAATTTTTGTA GAGACAAGGTTTTGCCATGTTGTCCAGGCTGGTCTTGAACTCCTGGGCTC AAGGGATCCGGCCACCTCAGCCTCCCAAAGTGCTAGGATTATAGGCATGA GCCACTGTGCCCAGCCTACCTTCAACGTATCTAACTGGTTACTAACTTTT AGGATTCGGCCTATGTCTCACAACCTTCTTGCTTACTCAACATCCTTGTC TCTTAAGCCACTAGCTTCTTCTCTATGGTTAACACTTTTTATGAGTTTTA TTCATCTGCTTATTTTTCTTATCCTCTATACCAGAATTGAATATTTTCAA ATAAAGCACACTCATGTTACAATCTTTGAAATGAAAAAAAAAAATGCATA GGATTAGAAAAGAAACCAATTTTAATAAACTATATTTTGAAGTATAGTTC TATATTAAACAACAAGATCTAGGCCAGGTGCAGTGGCTCATGCCTGTAAT CCCAGCAATTTGGGAAGTCGAGGTGGGAGGATTGCTTGAGGCCAGGGGTT CAAGACCAGCCTGGGCAACATGGAGAGATTCCCCATCTCTTTCTTTACAC ACACACACACACACACACACAAAATATCTGATAGCAACAGGTGCAGTCAT TACCACAATTTCGAGTAGTGATGAGCTTAATAATATTTCGAGTTATCACC AACAACTGTAAAGTAACATGAAAACGTCTGTGATGACTATTGCCCACAAA GTCACAGGTACTGCTAATACTCCTGGTATTTGTAGTCAAATTCATAATAA AGGAAATGCTAGGTTTCAGTTGGTATTTTGTCCCGACGGTCTGTGGACGG CAGGTTAGAACGCCCGTCCAAGCCAGGAGGGTGGACCTAGCACTGCAGGG TCCACCTCGGGCCAATCAACTATATTCCCGAGGCGGGGGCCTGCGCTTCC CGGACCCAGCTGCCCTCAGGGGAGAGAGGACACACTTAAGAGTTTGGGGC CGGCGTGGTAGCTCATGCCCCTGATCCCAGCACTTCGGGAGGCTGAGGCG TGAAGATCACTTGTAGCAGGAGTTTGAGACCAGTCTAGCCAACTTGGCGA GACCCTGTCCCTAAAAAAAATTTTTTTTTAATTAGCCAGTTGTGGTGAGC GCCTGTAGTCCCAGCTACTCGGGAGGCTGAGGTGGGAGGATCGCTGGGCT CAGGAGTTCCAGACTGCAGTGAGCCATGATGGCGGCACTGCACTCCAGCG CGGTGAGACTCAGTCTCAAAAATAAAAGGGGGAGGGGTTGGGGGTAAAAT TAGTTGTGAAATCAAGTAAGACTTCCTGGGACAGAACAATCAAAGGGGTG GCGCCGGGTCCTCCAAAGAGCTACTAGCTCAGCCCAAGCCCCGCCTCGGC CCCCAGGGCAGCGGCCCGCAGAGCTCCACCCGGCAGGCGCCCGGGAAACT CCGCCCCCCGGCCGGCAGGGGGCGCGCGCGCCGCCGGCCCCGCCCCGTGG ACGCGGGTTCCGTGGGCGTTCCCGCGGCCAGGCATCAGCAATCTATCAGG GAACGGCGGTGGCCGGTGCGGCGTGTTCGGTGGCGGCTCTGGCCGCTCAG GCGCCTGCGGCTGGGTGAGCGCACGCGAGGCGGCGAGGCGGCAGCGTGTT TCTAGGTCGTGGCGTCGGGCTTCCGGAGCTTTGGCGGCAGCTAGGGGAGG ATG

TNFα

Tumor necrosis factor is a cytokine produced primarily by activated macrophages (Ml type) and other cells including CD4+ lymphocytes, NK cells and neurons (Pfeffer K. 2003 Cytokine Growth Factor Rev. 14(3-4):185-91) to regulate immune cells during an acute inflammatory response. TNF was originally characterized its ability to induce tumor cell apoptosis and cachexia, however, its roles are now recognized to impart both beneficial (inflammation and in protective immune responses against a variety of infectious pathogens) and detrimental effects (sepsis, cancer, autoimmune disease). TNF, an endogenous pyrogen, induces fever, apoptotic cell death, cachexia, inflammation, inhibits tumorigenesis and viral replication and mediates sepsis by responding to IL-1 and IL-6 producing cells. Dysregulation of TNF production has been implicated in a variety of human diseases including Alzheimer's disease, cancer, major depression and inflammatory bowel disease (IBD). TNFα can be produced ectopically in the setting of malignancy and parallels parathyroid hormone both in causing secondary hypercalcemia and in the cancers with which excessive production is associated.

Protein: TNFα Gene: TNFα (Homo sapiens, chromosome 6, 31543344-31546113 [NCBI Reference Sequence: NC_(—)000006.11]; start site location: 31543519; strand: positive)

Gene Identification GeneID 7124 HGNC 11892 HPRD 01855 MIM 191160

Targeted Sequences Relative upstream location De- to gene Sequence sign start ID No: ID Sequence (5′-3′) site 10095 CGGGGAAAGAATCATTCAACCAGCGG 229 10096 TNF1 CGGTTTCTTCTCCATCGCGGGGGCG 326 10129 CTGCTCCGATTCCGAGGGGGGTCTTCT 412 10154 CTCCGTGTGGGGCTCTGGTCGGCAGCT 1464 10207 CGCAGCCCCGTGGTACATCGAGTGCAGC 2151

Target Shift Sequences Relative upstream location to Sequence gene start ID No: Sequence (5′-3′) site 10095 CGGGGAAAGAATCATTCAACCAGCGG 229 10096 CGGTTTCTTCTCCATCGCGGGGGCG 326 10097 GGTTTCTTCTCCATCGCGGG 327 10098 GTTTCTTCTCCATCGCGGGG 328 10099 TTTCTTCTCCATCGCGGGGG 329 10100 TTCTTCTCCATCGCGGGGGC 330 10101 TCTTCTCCATCGCGGGGGCG 331 10102 CTTCTCCATCGCGGGGGCGG 332 10103 TTCTCCATCGCGGGGGCGGG 333 10104 TCTCCATCGCGGGGGCGGGG 334 10105 CTCCATCGCGGGGGCGGGGA 335 10106 TCCATCGCGGGGGCGGGGAT 336 10107 CCATCGCGGGGGCGGGGATT 337 10108 CATCGCGGGGGCGGGGATTT 338 10109 ATCGCGGGGGCGGGGATTTG 339 10110 TCGCGGGGGCGGGGATTTGG 340 10111 TCGGTTTCTTCTCCATCGCG 325 10112 CTCGGTTTCTTCTCCATCGC 324 10113 TCTCGGTTTCTTCTCCATCG 323 10114 GTCTCGGTTTCTTCTCCATC 322 10115 TGTCTCGGTTTCTTCTCCAT 321 10116 CTGTCTCGGTTTCTTCTCCA 320 10117 TCTGTCTCGGTTTCTTCTCC 319 10118 TTCTGTCTCGGTTTCTTCTC 318 10119 CTTCTGTCTCGGTTTCTTCT 317 10120 CCTTCTGTCTCGGTTTCTTC 316 10121 ACCTTCTGTCTCGGTTTCTT 315 10122 CACCTTCTGTCTCGGTTTCT 314 10123 GCACCTTCTGTCTCGGTTTC 313 10124 TGCACCTTCTGTCTCGGTTT 312 10125 CTGCACCTTCTGTCTCGGTT 311 10126 CCTGCACCTTCTGTCTCGGT 310 10127 CCCTGCACCTTCTGTCTCGG 309 10128 GCCCTGCACCTTCTGTCTCG 308 10129 CTGCTCCGATTCCGAGGGGGGTCTTCT 412 10130 TGCTCCGATTCCGAGGGGGG 413 10131 GCTCCGATTCCGAGGGGGGT 414 10132 CTCCGATTCCGAGGGGGGTC 415 10133 TCCGATTCCGAGGGGGGTCT 416 10134 CCGATTCCGAGGGGGGTCTT 417 10135 CGATTCCGAGGGGGGTCTTC 418 10136 GATTCCGAGGGGGGTCTTCT 419 10137 ATTCCGAGGGGGGTCTTCTG 420 10138 TTCCGAGGGGGGTCTTCTGG 421 10139 TCCGAGGGGGGTCTTCTGGG 422 10140 CCGAGGGGGGTCTTCTGGGC 423 10141 CGAGGGGGGTCTTCTGGGCC 424 10142 CCTGCTCCGATTCCGAGGGG 411 10143 CCCTGCTCCGATTCCGAGGG 410 10144 TCCCTGCTCCGATTCCGAGG 409 10145 CTCCCTGCTCCGATTCCGAG 408 10146 CCTCCCTGCTCCGATTCCGA 407 10147 TCCTCCCTGCTCCGATTCCG 406 10148 ATCCTCCCTGCTCCGATTCC 405 10149 CATCCTCCCTGCTCCGATTC 404 10150 CCATCCTCCCTGCTCCGATT 403 10151 CCCATCCTCCCTGCTCCGAT 402 10152 CCCCATCCTCCCTGCTCCGA 401 10153 TCCCCATCCTCCCTGCTCCG 400 10154 CTCCGTGTGGGGCTCTGGTCGGCAGCT 1464 10155 TCCGTGTGGGGCTCTGGTCG 1465 10156 CCGTGTGGGGCTCTGGTCGG 1466 10157 CGTGTGGGGCTCTGGTCGGC 1467 10158 GTGTGGGGCTCTGGTCGGCA 1468 10159 TGTGGGGCTCTGGTCGGCAG 1469 10160 GTGGGGCTCTGGTCGGCAGC 1470 10161 TGGGGCTCTGGTCGGCAGCT 1471 10162 GGGGCTCTGGTCGGCAGCTG 1472 10163 GGGCTCTGGTCGGCAGCTGG 1473 10164 GGCTCTGGTCGGCAGCTGGC 1474 10165 GCTCTGGTCGGCAGCTGGCT 1475 10166 CTCTGGTCGGCAGCTGGCTT 1476 10167 TCTGGTCGGCAGCTGGCTTT 1477 10168 CTGGTCGGCAGCTGGCTTTC 1478 10169 TGGTCGGCAGCTGGCTTTCA 1479 10170 GGTCGGCAGCTGGCTTTCAG 1480 10171 GTCGGCAGCTGGCTTTCAGA 1481 10172 TCGGCAGCTGGCTTTCAGAG 1482 10173 CGGCAGCTGGCTTTCAGAGC 1483 10174 CCTCCGTGTGGGGCTCTGGT 1463 10175 GCCTCCGTGTGGGGCTCTGG 1462 10176 TGCCTCCGTGTGGGGCTCTG 1461 10177 ATGCCTCCGTGTGGGGCTCT 1460 10178 GATGCCTCCGTGTGGGGCTC 1459 10179 AGATGCCTCCGTGTGGGGCT 1458 10180 CAGATGCCTCCGTGTGGGGC 1457 10181 GCAGATGCCTCCGTGTGGGG 1456 10182 TGCAGATGCCTCCGTGTGGG 1455 10183 GTGCAGATGCCTCCGTGTGG 1454 10184 GGTGCAGATGCCTCCGTGTG 1453 10185 GGGTGCAGATGCCTCCGTGT 1452 10186 AGGGTGCAGATGCCTCCGTG 1451 10187 GAGGGTGCAGATGCCTCCGT 1450 10188 CGAGGGTGCAGATGCCTCCG 1449 10189 TCGAGGGTGCAGATGCCTCC 1448 10190 ATCGAGGGTGCAGATGCCTC 1447 10191 CATCGAGGGTGCAGATGCCT 1446 10192 TCATCGAGGGTGCAGATGCC 1445 10193 TTCATCGAGGGTGCAGATGC 1444 10194 CTTCATCGAGGGTGCAGATG 1443 10195 GCTTCATCGAGGGTGCAGAT 1442 10196 GGCTTCATCGAGGGTGCAGA 1441 10197 GGGCTTCATCGAGGGTGCAG 1440 10198 TGGGCTTCATCGAGGGTGCA 1439 10199 TTGGGCTTCATCGAGGGTGC 1438 10200 ATTGGGCTTCATCGAGGGTG 1437 10201 TATTGGGCTTCATCGAGGGT 1436 10202 TTATTGGGCTTCATCGAGGG 1435 10203 TTTATTGGGCTTCATCGAGG 1434 10204 GTTTATTGGGCTTCATCGAG 1433 10205 GGTTTATTGGGCTTCATCGA 1432 10206 AGGTTTATTGGGCTTCATCG 1431 10207 CGCAGCCCCGTGGTACATCGAGTGCAGC 2151 10208 GCAGCCCCGTGGTACATCGA 2152 10209 CAGCCCCGTGGTACATCGAG 2153 10210 AGCCCCGTGGTACATCGAGT 2154 10211 GCCCCGTGGTACATCGAGTG 2155 10212 CCCCGTGGTACATCGAGTGC 2156 10213 CCCGTGGTACATCGAGTGCA 2157 10214 CCGTGGTACATCGAGTGCAG 2158 10215 CGTGGTACATCGAGTGCAGC 2159 10216 GTGGTACATCGAGTGCAGCC 2160 10217 TGGTACATCGAGTGCAGCCA 2161 10218 GGTACATCGAGTGCAGCCAG 2162 10219 GTACATCGAGTGCAGCCAGG 2163 10220 TACATCGAGTGCAGCCAGGG 2164 10221 ACATCGAGTGCAGCCAGGGT 2165 10222 CATCGAGTGCAGCCAGGGTT 2166 10223 ATCGAGTGCAGCCAGGGTTC 2167 10224 TCGAGTGCAGCCAGGGTTCC 2168 10225 CGAGTGCAGCCAGGGTTCCT 2169 10226 ACGCAGCCCCGTGGTACATC 2150 10227 AACGCAGCCCCGTGGTACAT 2149 10228 GAACGCAGCCCCGTGGTACA 2148 10229 GGAACGCAGCCCCGTGGTAC 2147 10230 TGGAACGCAGCCCCGTGGTA 2146 10231 CTGGAACGCAGCCCCGTGGT 2145 10232 GCTGGAACGCAGCCCCGTGG 2144 10233 AGCTGGAACGCAGCCCCGTG 2143 10234 GAGCTGGAACGCAGCCCCGT 2142 10235 TGAGCTGGAACGCAGCCCCG 2141 10236 GTGAGCTGGAACGCAGCCCC 2140 10237 GGTGAGCTGGAACGCAGCCC 2139 10238 GGGTGAGCTGGAACGCAGCC 2138 10239 TGGGTGAGCTGGAACGCAGC 2137 10240 CTGGGTGAGCTGGAACGCAG 2136 10241 CCTGGGTGAGCTGGAACGCA 2135 10242 CCCTGGGTGAGCTGGAACGC 2134 10243 TCCCTGGGTGAGCTGGAACG 2133

Hot Zones (Relative upstream location to gene start site) 168-450 1430-1520 2150-2240

Examples

In FIG. 62, In MCF7 (human mammary breast cell line), TNF1 (312) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The TNFα sequence TNF1 (312) fits the independent and dependent DNAi motif claims.

The secondary structure for TNF1 (312) is shown in FIG. 63.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11982) CTCACTGTCTCTCTCTCTCTCTCTCTTTCTCTGCAGGTTCTCCCCATGAC ACCACCTGAACGTCTCTTCCTCCCAAGGGTGTGTGGCACCACCCTACACC TCCTCCTTCTGGGGCTGCTGCTGGTTCTGCTGCCTGGGGCCCAGGTGAGG CAGCAGGAGAATGGGGGCTGCTGGGGTGGCTCAGCCAAACCTTGAGCCCT AGAGCCCCCCTCAACTCTGTTCTCCCCTAGGGGCTCCCTGGTGTTGGCCT CACACCTTCAGCTGCCCAGACTGCCCGTCAGCACCCCAAGATGCATCTTG CCCACAGCACCCTCAAACCTGCTGCTCACCTCATTGGTAAACATCCACCT GACCTCCCAGACATGTCCCCACCAGCTCTCCTCCTACCCCTGCCTCAGGA ACCCAAGCATCCACCCCTCTCCCCCAACTTCCCCCACGCTAAAAAAAACA GAGGGAGCCCACTCCTATGCCTCCCCCTGCCATCCCCCAGGAACTCAGTT GTTCAGTGCCCACTTCCTCAGGGATTGAGACCTCTGATCCAGACCCCTGA TCTCCCACCCCCATCCCCTATGGCTCTTCCTAGGAGACCCCAGCAAGCAG AACTCACTGCTCTGGAGAGCAAACACGGACCGTGCCTTCCTCCAGGATGG TTTCTCCTTGAGCAACAATTCTCTCCTGGTCCCCACCAGTGGCATCTACT TCGTCTACTCCCAGGTGGTCTTCTCTGGGAAAGCCTACTCTCCCAAGGCC ACCTCCTCCCCACTCTACCTGGCCCATGAGGTCCAGCTCTTCTCCTCCCA GTACCCCTTCCATGTGCCTCTCCTCAGCTCCCAGAAGATGGTGTATCCAG GGCTGCAGGAACCCTGGCTGCACTCGATGTACCACGGGGCTGCGTTCCAG CTCACCCAGGGAGACCAGCTATCCACCCACACAGATGGCATCCCCCACCT AGTCCTCAGCCCTAGTACTGTCTTCTTTGGAGCCTTCGCTCTGTAGAACT TGGAAAAATCCAGAAAGAAAAAATAATTGATTTCAAGACCTTCTCCCCAT TCTGCCTCCATTCTGACCATTTCAGGGGTCGTCACCACCTCTCCTTTGGC CATTCCAACAGCTCAAGTCTTCCCTGATCAAGTCACCGGAGCTTTCAAAG AAGGAATTCTAGGCATCCCAGGGGACCACACCTCCCTGAACCATCCCTGA TGTCTGTCTGGCTGAGGATTTCAAGCCTGCCTAGGAATTCCCAGCCCAAA GCTGTTGGTCTGTCCCACCAGCTAGGTGGGGCCTAGATCCACACACAGAG GAAGAGCAGGCACATGGAGGAGCTTGGGGGATGACTAGAGGCAGGGAGGG GACTATTTATGAAGGCAAAAAAATTAAATTATTTATTTATGGAGGATGGA GAGAGGGGAATAATAGAAGAACATCCAAGGAGAAACAGAGACAGGCCCAA GAGATGAAGAGTGAGAGGGCATGCGCACAAGGCTGACCAAGAGAGAAAGA AGTAGGCATGAGGGATCACAGGGCCCCAGAAGGCAGGGAAAGGCTCTGAA AGCCAGCTGCCGACCAGAGCCCCACACGGAGGCATCTGCACCCTCGATGA AGCCCAATAAACCTCTTTTCTCTGAAATGCTGTCTGCTTGTGTGTGTGTG TCTGGGAGTGAGAACTTCCCAGTCTATCTAAGGAATGGAGGGAGGGACAG AGGGCTCAAAGGGAGCAAGAGCTGTGGGGAGAACAAAAGGATAAGGGCTC AGAGAGCTTCAGGGATATGTGATGGACTCACCAGGTGAGGCCGCCAGACT GCTGCAGGGGAAGCAAAGGAGAAGCTGAGAAGATGAAGGAAAAGTCAGGG TCTGGAGGGGCGGGGGTCAGGGAGCTCCTGGGAGATATGGCCACATGTAG CGGCTCTGAGGAATGGGTTACAGGAGACCTCTGGGGAGATGTGACCACAG CAATGGGTAGGAGAATGTCCAGGGCTATGGAAGTCGAGTATGGGGACCCC CCCTTAACGAAGACAGGGCCATGTAGAGGGCCCCAGGGAGTGAAAGAGCC TCCAGGACCTCCAGGTATGGAATACAGGGGACGTTTAAGAAGATATGGCC ACACACTGGGGCCCTGAGAAGTGAGAGCTTCATGAAAAAAATCAGGGACC CCAGAGTTCCTTGGAAGCCAAGACTGAAACCAGCATTATGAGTCTCCGGG TCAGAATGAAAGAAGAAGGCCTGCCCCAGTGGGGTCTGTGAATTCCCGGG GGTGATTTCACTCCCCGGGGCTGTCCCAGGCTTGTCCCTGCTACCCCCAC CCAGCCTTTCCTGAGGCCTCAAGCCTGCCACCAAGCCCCCAGCTCCTTCT CCCCGCAGGGACCCAAACACAGGCCTCAGGACTCAACACAGCTTTTCCCT CCAACCCCGTTTTCTCTCCCTCAAGGACTCAGCTTTCTGAAGCCCCTCCC AGTTCTAGTTCTATCTTTTTCCTGCATCCTGTCTGGAAGTTAGAAGGAAA CAGACCACAGACCTGGTCCCCAAAAGAAATGGAGGCAATAGGTTTTGAGG GGCATGGGGACGGGGTTCAGCCTCCAGGGTCCTACACACAAATCAGTCAG TGGCCCAGAAGACCCCCCTCGGAATCGGAGCAGGGAGGATGGGGAGTGTG AGGGGTATCCTTGATGCTTGTGTGTCCCCAACTTTCCAAATCCCCGCCCC CGCGATGGAGAAGAAACCGAGACAGAAGGTGCAGGGCCCACTACCGCTTC CTCCAGATGAGCTCATGGGTTTCTCCACCAAGGAAGTTTTCCGCTGGTTG AATGATTCTTTCCCCGCCCTCCTCTCGCCCCAGGGACATATAAAGGCAGT TGTTGGCACACCCAGCCAGCAGACGCTCCCTCAGCAAGGACAGCAGAGGA CCAGCTAAGAGGGAGAGAAGCAACTACAGACCCCCCCTGAAAACAACCCT CAGACGCCACATCCCCTGACAAGCTGCCAGGCAGGTTCTCTTCCTCTCAC ATACTGACCCACGGCTCCACCCTCTCTCCCCTGGAAAGGACACC ATG

ITGA4

Integrins are ubiquitously expressed adhesion molecules. They are cell-surface receptors that exist as heterodimers of alpha and beta subunits. ITGA4 encodes an alpha 4 chain. Unlike other integrin alpha chains, alpha 4 neither contains an I-domain, nor undergoes disulfide-linked cleavage. Alpha 4 chain associates with either beta 1 chain or beta 7 chain. At physiological conditions, integrins are highly glycosylated and contain a Ca2+ or Mg2+ ion, which is essential for ligand binding. Integrin receptors are critical for cell attachment to the extracellular matrix (ECM) and this is mediated through integrin-fibronectin, -vitronectin, -collagen and -laminin interactions. Intracellularly, integrins form adhesion complexes with proteins including talin, vinculin, paxillin and alpha-actinin. They also regulate kinases, such as focal adhesion kinase and Src family kinases, to mediate attachment to the actin cytoskeleton. Integrins also have a significant role in cell signaling and can activate protein kinases involved in the regulation of cell growth, division, survival, differentiation, migration and apoptosis. Glycoprotein II/IIIb (alphaIIbeta3) is an integrin receptor found on the surface of platelets. It is involved in the cross-linking of platelets with fibrin, and so has a vital role in blood clot formation.

Protein: ITGA4 Gene: ITGA4 (CD49D) (Homo sapiens, chromosome 2, 182321619-182402474 [NCBI Reference Sequence: NC_(—)000002.11]; start site location: 182322383; strand: positive)

Gene Identification GeneID 3676 HGNC 6140 MIM 192975

Targeted Sequences Relative upstream location Se- to gene quence Design start ID No: ID Sequence (5′-3′) site 10244 GCGCTCTCGGTGGGGAACATTCAACAC 1 10252 CGGGATGCGACGGTTGGCCAACGG 54 10278 CGCAGCGTGTCCGGCGCCAGCGGGC 102 10299 CGGCCCACCGCGGGCGGAGCGTTCG 160 10449 CGCGCACTCGCCCGGCCCCACTCCCG 201 10599 CGCCAGCCGGGAGCTTCGGGTGCTCGCG 235 10749 CGGGTACGGGCCGCTGGGTGGGGTCCCG 272 10899 GTGCGGAGGCGCAGGGCCGGGCTCCG 306 10900 CTACGCGCGGCTGCAGGGGGCGC 339 10938 CTGCGCAGGACTCGCGTCCTGGCCCG 375 11009 CCCGCAGAGCGCGGGATGGCTC 411 11080 CGGACCTGATGGGGCACGGGCTTCCCC 448 11117 CGGTGGTTGGGGCCTAGAAGCG 481 11154 CGCGCCCCTCGCTGTGACCGCCCAGCCCG 524 11203 CGGGGAGTGGGACTGCGGCGGGGAGCCG 580 11208 ACTCGCCGAAGGCCCCTGGGGAAC 718 11222 CGGGCTGCATGCGTGAGCAGG 840 11252 CGGCAGGCGGTTTAGGCTGTGGCTG 885 11278 CCGATTCGGATTGCTCCAGCTGG 962 11289 CGCACCCACTCAGTTGCCACGGG 1008 11327 CGGAGACCCACAACGCAACACACC 1099

Target Shift Sequences Relative upstream location to Sequence gene start ID No: Sequence (5′-3′) site 10244 GCGCTCTCGGTGGGGAACATTCAACAC 1 10245 CGCTCTCGGTGGGGAACATT 2 10246 GCTCTCGGTGGGGAACATTC 3 10247 CTCTCGGTGGGGAACATTCA 4 10248 TCTCGGTGGGGAACATTCAA 5 10249 CTCGGTGGGGAACATTCAAC 6 10250 TCGGTGGGGAACATTCAACA 7 10251 CGGTGGGGAACATTCAACAC 8 10252 CGGGATGCGACGGTTGGCCAACGG 54 10253 GGGATGCGACGGTTGGCCAA 55 10254 GGATGCGACGGTTGGCCAAC 56 10255 GATGCGACGGTTGGCCAACG 57 10256 ATGCGACGGTTGGCCAACGG 58 10257 TGCGACGGTTGGCCAACGGG 59 10258 GCGACGGTTGGCCAACGGGG 60 10259 CGACGGTTGGCCAACGGGGA 61 10260 ACGGGATGCGACGGTTGGCC 53 10261 CACGGGATGCGACGGTTGGC 52 10262 GCACGGGATGCGACGGTTGG 51 10263 TGCACGGGATGCGACGGTTG 50 10264 TTGCACGGGATGCGACGGTT 49 10265 GTTGCACGGGATGCGACGGT 48 10266 AGTTGCACGGGATGCGACGG 47 10267 AAGTTGCACGGGATGCGACG 46 10268 AAAGTTGCACGGGATGCGAC 45 10269 CAAAGTTGCACGGGATGCGA 44 10270 CCAAAGTTGCACGGGATGCG 43 10271 CCCAAAGTTGCACGGGATGC 42 10272 CCCCAAAGTTGCACGGGATG 41 10273 ACCCCAAAGTTGCACGGGAT 40 10274 TACCCCAAAGTTGCACGGGA 39 10275 CTACCCCAAAGTTGCACGGG 38 10276 ACTACCCCAAAGTTGCACGG 37 10277 CACTACCCCAAAGTTGCACG 36 10278 CGCAGCGTGTCCGGCGCCAGCGGGC 102 10279 GCAGCGTGTCCGGCGCCAGC 103 10280 CAGCGTGTCCGGCGCCAGCG 104 10281 AGCGTGTCCGGCGCCAGCGG 105 10282 GCGTGTCCGGCGCCAGCGGG 106 10283 CGTGTCCGGCGCCAGCGGGC 107 10284 GTGTCCGGCGCCAGCGGGCT 108 10285 TGTCCGGCGCCAGCGGGCTA 109 10286 GTCCGGCGCCAGCGGGCTAA 110 10287 TCCGGCGCCAGCGGGCTAAA 111 10288 CCGGCGCCAGCGGGCTAAAG 112 10289 CGGCGCCAGCGGGCTAAAGG 113 10290 GCGCAGCGTGTCCGGCGCCA 101 10291 GGCGCAGCGTGTCCGGCGCC 100 10292 AGGCGCAGCGTGTCCGGCGC 99 10293 GAGGCGCAGCGTGTCCGGCG 98 10294 TGAGGCGCAGCGTGTCCGGC 97 10295 ATGAGGCGCAGCGTGTCCGG 96 10296 GATGAGGCGCAGCGTGTCCG 95 10297 AGATGAGGCGCAGCGTGTCC 94 10298 GAGATGAGGCGCAGCGTGTC 93 10299 CGGCCCACCGCGGGCGGAGCGTTCG 160 10300 GGCCCACCGCGGGCGGAGCG 161 10301 GCCCACCGCGGGCGGAGCGT 162 10302 CCCACCGCGGGCGGAGCGTT 163 10303 CCACCGCGGGCGGAGCGTTC 164 10304 CACCGCGGGCGGAGCGTTCG 165 10305 ACCGCGGGCGGAGCGTTCGG 166 10306 CCGCGGGCGGAGCGTTCGGG 167 10307 CGCGGGCGGAGCGTTCGGGC 168 10308 GCGGGCGGAGCGTTCGGGCC 169 10309 CGGGCGGAGCGTTCGGGCCG 170 10310 GGGCGGAGCGTTCGGGCCGG 171 10311 GGCGGAGCGTTCGGGCCGGC 172 10312 GCGGAGCGTTCGGGCCGGCC 173 10313 CGGAGCGTTCGGGCCGGCCT 174 10314 GGAGCGTTCGGGCCGGCCTG 175 10315 GAGCGTTCGGGCCGGCCTGG 176 10316 AGCGTTCGGGCCGGCCTGGG 177 10317 GCGTTCGGGCCGGCCTGGGA 178 10318 CGTTCGGGCCGGCCTGGGAT 179 10319 GTTCGGGCCGGCCTGGGATG 180 10320 TTCGGGCCGGCCTGGGATGC 181 10321 TCGGGCCGGCCTGGGATGCC 182 10322 CGGGCCGGCCTGGGATGCCG 183 10323 GGGCCGGCCTGGGATGCCGC 184 10324 GGCCGGCCTGGGATGCCGCG 185 10325 GCCGGCCTGGGATGCCGCGC 186 10326 CCGGCCTGGGATGCCGCGCA 187 10327 CGGCCTGGGATGCCGCGCAC 188 10328 GGCCTGGGATGCCGCGCACT 189 10329 GCCTGGGATGCCGCGCACTC 190 10330 CCTGGGATGCCGCGCACTCG 191 10331 CTGGGATGCCGCGCACTCGC 192 10332 TGGGATGCCGCGCACTCGCC 193 10333 GGGATGCCGCGCACTCGCCC 194 10334 GGATGCCGCGCACTCGCCCG 195 10335 GATGCCGCGCACTCGCCCGG 196 10336 ATGCCGCGCACTCGCCCGGC 197 10337 TGCCGCGCACTCGCCCGGCC 198 10338 GCCGCGCACTCGCCCGGCCC 199 10339 CCGCGCACTCGCCCGGCCCC 200 10340 CGCGCACTCGCCCGGCCCCA 201 10341 GCGCACTCGCCCGGCCCCAC 202 10342 CGCACTCGCCCGGCCCCACT 203 10343 GCACTCGCCCGGCCCCACTC 204 10344 CACTCGCCCGGCCCCACTCC 205 10345 ACTCGCCCGGCCCCACTCCC 206 10346 CTCGCCCGGCCCCACTCCCG 207 10347 TCGCCCGGCCCCACTCCCGG 208 10348 CGCCCGGCCCCACTCCCGGT 209 10349 GCCCGGCCCCACTCCCGGTT 210 10350 CCCGGCCCCACTCCCGGTTT 211 10351 CCGGCCCCACTCCCGGTTTC 212 10352 CGGCCCCACTCCCGGTTTCT 213 10353 GGCCCCACTCCCGGTTTCTG 214 10354 GCCCCACTCCCGGTTTCTGC 215 10355 CCCCACTCCCGGTTTCTGCC 216 10356 CCCACTCCCGGTTTCTGCCG 217 10357 CCACTCCCGGTTTCTGCCGC 218 10358 CACTCCCGGTTTCTGCCGCC 219 10359 ACTCCCGGTTTCTGCCGCCA 220 10360 CTCCCGGTTTCTGCCGCCAG 221 10361 TCCCGGTTTCTGCCGCCAGC 222 10362 CCCGGTTTCTGCCGCCAGCC 223 10363 CCGGTTTCTGCCGCCAGCCG 224 10364 CGGTTTCTGCCGCCAGCCGG 225 10365 GGTTTCTGCCGCCAGCCGGG 226 10366 GTTTCTGCCGCCAGCCGGGA 227 10367 TTTCTGCCGCCAGCCGGGAG 228 10368 TTCTGCCGCCAGCCGGGAGC 229 10369 TCTGCCGCCAGCCGGGAGCT 230 10370 CTGCCGCCAGCCGGGAGCTT 231 10371 TGCCGCCAGCCGGGAGCTTC 232 10372 GCCGCCAGCCGGGAGCTTCG 233 10373 CCGCCAGCCGGGAGCTTCGG 234 10374 CGCCAGCCGGGAGCTTCGGG 235 10375 GCCAGCCGGGAGCTTCGGGT 236 10376 CCAGCCGGGAGCTTCGGGTG 237 10377 CAGCCGGGAGCTTCGGGTGC 238 10378 AGCCGGGAGCTTCGGGTGCT 239 10379 GCCGGGAGCTTCGGGTGCTC 240 10380 CCGGGAGCTTCGGGTGCTCG 241 10381 CGGGAGCTTCGGGTGCTCGC 242 10382 GGGAGCTTCGGGTGCTCGCG 243 10383 GGAGCTTCGGGTGCTCGCGC 244 10384 GAGCTTCGGGTGCTCGCGCT 245 10385 AGCTTCGGGTGCTCGCGCTG 246 10386 GCTTCGGGTGCTCGCGCTGC 247 10387 CTTCGGGTGCTCGCGCTGCT 248 10388 TTCGGGTGCTCGCGCTGCTT 249 10389 TCGGGTGCTCGCGCTGCTTC 250 10390 CGGGTGCTCGCGCTGCTTCT 251 10391 GGGTGCTCGCGCTGCTTCTC 252 10392 GGTGCTCGCGCTGCTTCTCC 253 10393 GTGCTCGCGCTGCTTCTCCG 254 10394 TGCTCGCGCTGCTTCTCCGG 255 10395 GCTCGCGCTGCTTCTCCGGG 256 10396 CTCGCGCTGCTTCTCCGGGT 257 10397 TCGCGCTGCTTCTCCGGGTA 258 10398 CGCGCTGCTTCTCCGGGTAC 259 10399 GCGCTGCTTCTCCGGGTACG 260 10400 CGCTGCTTCTCCGGGTACGG 261 10401 GCTGCTTCTCCGGGTACGGG 262 10402 CTGCTTCTCCGGGTACGGGC 263 10403 TGCTTCTCCGGGTACGGGCC 264 10404 GCTTCTCCGGGTACGGGCCG 265 10405 CTTCTCCGGGTACGGGCCGC 266 10406 TTCTCCGGGTACGGGCCGCT 267 10407 TCTCCGGGTACGGGCCGCTG 268 10408 CTCCGGGTACGGGCCGCTGG 269 10409 TCCGGGTACGGGCCGCTGGG 270 10410 CCGGGTACGGGCCGCTGGGT 271 10411 CGGGTACGGGCCGCTGGGTG 272 10412 GGGTACGGGCCGCTGGGTGG 273 10413 GGTACGGGCCGCTGGGTGGG 274 10414 GTACGGGCCGCTGGGTGGGG 275 10415 TACGGGCCGCTGGGTGGGGT 276 10416 ACGGGCCGCTGGGTGGGGTC 277 10417 CGGGCCGCTGGGTGGGGTCC 278 10418 GGGCCGCTGGGTGGGGTCCC 279 10419 GGCCGCTGGGTGGGGTCCCG 280 10420 GCCGCTGGGTGGGGTCCCGG 281 10421 CCGCTGGGTGGGGTCCCGGG 282 10422 CGCTGGGTGGGGTCCCGGGC 283 10423 GCTGGGTGGGGTCCCGGGCG 284 10424 CTGGGTGGGGTCCCGGGCGT 285 10425 TGGGTGGGGTCCCGGGCGTG 286 10426 GGGTGGGGTCCCGGGCGTGG 287 10427 GGTGGGGTCCCGGGCGTGGT 288 10428 GTGGGGTCCCGGGCGTGGTG 289 10429 TGGGGTCCCGGGCGTGGTGC 290 10430 GGGGTCCCGGGCGTGGTGCG 291 10431 GGGTCCCGGGCGTGGTGCGG 292 10432 GGTCCCGGGCGTGGTGCGGA 293 10433 GTCCCGGGCGTGGTGCGGAG 294 10434 TCCCGGGCGTGGTGCGGAGG 295 10435 CCCGGGCGTGGTGCGGAGGC 296 10436 CCGGGCGTGGTGCGGAGGCG 297 10437 CGGGCGTGGTGCGGAGGCGC 298 10438 TCGGCCCACCGCGGGCGGAG 159 10439 GTCGGCCCACCGCGGGCGGA 158 10440 AGTCGGCCCACCGCGGGCGG 157 10441 AAGTCGGCCCACCGCGGGCG 156 10442 GAAGTCGGCCCACCGCGGGC 155 10443 GGAAGTCGGCCCACCGCGGG 154 10444 GGGAAGTCGGCCCACCGCGG 153 10445 GGGGAAGTCGGCCCACCGCG 152 10446 AGGGGAAGTCGGCCCACCGC 151 10447 GAGGGGAAGTCGGCCCACCG 150 10448 GGAGGGGAAGTCGGCCCACC 149 10449 CGCGCACTCGCCCGGCCCCACTCCCG 201 10450 GCGCACTCGCCCGGCCCCAC 202 10451 CGCACTCGCCCGGCCCCACT 203 10452 GCACTCGCCCGGCCCCACTC 204 10453 CACTCGCCCGGCCCCACTCC 205 10454 ACTCGCCCGGCCCCACTCCC 206 10455 CTCGCCCGGCCCCACTCCCG 207 10456 TCGCCCGGCCCCACTCCCGG 208 10457 CGCCCGGCCCCACTCCCGGT 209 10458 GCCCGGCCCCACTCCCGGTT 210 10459 CCCGGCCCCACTCCCGGTTT 211 10460 CCGGCCCCACTCCCGGTTTC 212 10461 CGGCCCCACTCCCGGTTTCT 213 10462 GGCCCCACTCCCGGTTTCTG 214 10463 GCCCCACTCCCGGTTTCTGC 215 10464 CCCCACTCCCGGTTTCTGCC 216 10465 CCCACTCCCGGTTTCTGCCG 217 10466 CCACTCCCGGTTTCTGCCGC 218 10467 CACTCCCGGTTTCTGCCGCC 219 10468 ACTCCCGGTTTCTGCCGCCA 220 10469 CTCCCGGTTTCTGCCGCCAG 221 10470 TCCCGGTTTCTGCCGCCAGC 222 10471 CCCGGTTTCTGCCGCCAGCC 223 10472 CCGGTTTCTGCCGCCAGCCG 224 10473 CGGTTTCTGCCGCCAGCCGG 225 10474 GGTTTCTGCCGCCAGCCGGG 226 10475 GTTTCTGCCGCCAGCCGGGA 227 10476 TTTCTGCCGCCAGCCGGGAG 228 10477 TTCTGCCGCCAGCCGGGAGC 229 10478 TCTGCCGCCAGCCGGGAGCT 230 10479 CTGCCGCCAGCCGGGAGCTT 231 10480 TGCCGCCAGCCGGGAGCTTC 232 10481 GCCGCCAGCCGGGAGCTTCG 233 10482 CCGCCAGCCGGGAGCTTCGG 234 10483 CGCCAGCCGGGAGCTTCGGG 235 10484 GCCAGCCGGGAGCTTCGGGT 236 10485 CCAGCCGGGAGCTTCGGGTG 237 10486 CAGCCGGGAGCTTCGGGTGC 238 10487 AGCCGGGAGCTTCGGGTGCT 239 10488 GCCGGGAGCTTCGGGTGCTC 240 10489 CCGGGAGCTTCGGGTGCTCG 241 10490 CGGGAGCTTCGGGTGCTCGC 242 10491 GGGAGCTTCGGGTGCTCGCG 243 10492 GGAGCTTCGGGTGCTCGCGC 244 10493 GAGCTTCGGGTGCTCGCGCT 245 10494 AGCTTCGGGTGCTCGCGCTG 246 10495 GCTTCGGGTGCTCGCGCTGC 247 10496 CTTCGGGTGCTCGCGCTGCT 248 10497 TTCGGGTGCTCGCGCTGCTT 249 10498 TCGGGTGCTCGCGCTGCTTC 250 10499 CGGGTGCTCGCGCTGCTTCT 251 10500 GGGTGCTCGCGCTGCTTCTC 252 10501 GGTGCTCGCGCTGCTTCTCC 253 10502 GTGCTCGCGCTGCTTCTCCG 254 10503 TGCTCGCGCTGCTTCTCCGG 255 10504 GCTCGCGCTGCTTCTCCGGG 256 10505 CTCGCGCTGCTTCTCCGGGT 257 10506 TCGCGCTGCTTCTCCGGGTA 258 10507 CGCGCTGCTTCTCCGGGTAC 259 10508 GCGCTGCTTCTCCGGGTACG 260 10509 CGCTGCTTCTCCGGGTACGG 261 10510 GCTGCTTCTCCGGGTACGGG 262 10511 CTGCTTCTCCGGGTACGGGC 263 10512 TGCTTCTCCGGGTACGGGCC 264 10513 GCTTCTCCGGGTACGGGCCG 265 10514 CTTCTCCGGGTACGGGCCGC 266 10515 TTCTCCGGGTACGGGCCGCT 267 10516 TCTCCGGGTACGGGCCGCTG 268 10517 CTCCGGGTACGGGCCGCTGG 269 10518 TCCGGGTACGGGCCGCTGGG 270 10519 CCGGGTACGGGCCGCTGGGT 271 10520 CGGGTACGGGCCGCTGGGTG 272 10521 GGGTACGGGCCGCTGGGTGG 273 10522 GGTACGGGCCGCTGGGTGGG 274 10523 GTACGGGCCGCTGGGTGGGG 275 10524 TACGGGCCGCTGGGTGGGGT 276 10525 ACGGGCCGCTGGGTGGGGTC 277 10526 CGGGCCGCTGGGTGGGGTCC 278 10527 GGGCCGCTGGGTGGGGTCCC 279 10528 GGCCGCTGGGTGGGGTCCCG 280 10529 GCCGCTGGGTGGGGTCCCGG 281 10530 CCGCTGGGTGGGGTCCCGGG 282 10531 CGCTGGGTGGGGTCCCGGGC 283 10532 GCTGGGTGGGGTCCCGGGCG 284 10533 CTGGGTGGGGTCCCGGGCGT 285 10534 TGGGTGGGGTCCCGGGCGTG 286 10535 GGGTGGGGTCCCGGGCGTGG 287 10536 GGTGGGGTCCCGGGCGTGGT 288 10537 GTGGGGTCCCGGGCGTGGTG 289 10538 TGGGGTCCCGGGCGTGGTGC 290 10539 GGGGTCCCGGGCGTGGTGCG 291 10540 GGGTCCCGGGCGTGGTGCGG 292 10541 GGTCCCGGGCGTGGTGCGGA 293 10542 GTCCCGGGCGTGGTGCGGAG 294 10543 TCCCGGGCGTGGTGCGGAGG 295 10544 CCCGGGCGTGGTGCGGAGGC 296 10545 CCGGGCGTGGTGCGGAGGCG 297 10546 CGGGCGTGGTGCGGAGGCGC 298 10547 CCGCGCACTCGCCCGGCCCC 200 10548 GCCGCGCACTCGCCCGGCCC 199 10549 TGCCGCGCACTCGCCCGGCC 198 10550 ATGCCGCGCACTCGCCCGGC 197 10551 GATGCCGCGCACTCGCCCGG 196 10552 GGATGCCGCGCACTCGCCCG 195 10553 GGGATGCCGCGCACTCGCCC 194 10554 TGGGATGCCGCGCACTCGCC 193 10555 CTGGGATGCCGCGCACTCGC 192 10556 CCTGGGATGCCGCGCACTCG 191 10557 GCCTGGGATGCCGCGCACTC 190 10558 GGCCTGGGATGCCGCGCACT 189 10559 CGGCCTGGGATGCCGCGCAC 188 10560 CCGGCCTGGGATGCCGCGCA 187 10561 GCCGGCCTGGGATGCCGCGC 186 10562 GGCCGGCCTGGGATGCCGCG 185 10563 GGGCCGGCCTGGGATGCCGC 184 10564 CGGGCCGGCCTGGGATGCCG 183 10565 TCGGGCCGGCCTGGGATGCC 182 10566 TTCGGGCCGGCCTGGGATGC 181 10567 GTTCGGGCCGGCCTGGGATG 180 10568 CGTTCGGGCCGGCCTGGGAT 179 10569 GCGTTCGGGCCGGCCTGGGA 178 10570 AGCGTTCGGGCCGGCCTGGG 177 10571 GAGCGTTCGGGCCGGCCTGG 176 10572 GGAGCGTTCGGGCCGGCCTG 175 10573 CGGAGCGTTCGGGCCGGCCT 174 10574 GCGGAGCGTTCGGGCCGGCC 173 10575 GGCGGAGCGTTCGGGCCGGC 172 10576 GGGCGGAGCGTTCGGGCCGG 171 10577 CGGGCGGAGCGTTCGGGCCG 170 10578 GCGGGCGGAGCGTTCGGGCC 169 10579 CGCGGGCGGAGCGTTCGGGC 168 10580 CCGCGGGCGGAGCGTTCGGG 167 10581 ACCGCGGGCGGAGCGTTCGG 166 10582 CACCGCGGGCGGAGCGTTCG 165 10583 CCACCGCGGGCGGAGCGTTC 164 10584 CCCACCGCGGGCGGAGCGTT 163 10585 GCCCACCGCGGGCGGAGCGT 162 10586 GGCCCACCGCGGGCGGAGCG 161 10587 CGGCCCACCGCGGGCGGAGC 160 10588 TCGGCCCACCGCGGGCGGAG 159 10589 GTCGGCCCACCGCGGGCGGA 158 10590 AGTCGGCCCACCGCGGGCGG 157 10591 AAGTCGGCCCACCGCGGGCG 156 10592 GAAGTCGGCCCACCGCGGGC 155 10593 GGAAGTCGGCCCACCGCGGG 154 10594 GGGAAGTCGGCCCACCGCGG 153 10595 GGGGAAGTCGGCCCACCGCG 152 10596 AGGGGAAGTCGGCCCACCGC 151 10597 GAGGGGAAGTCGGCCCACCG 150 10598 GGAGGGGAAGTCGGCCCACC 149 10599 CGCCAGCCGGGAGCTTCGGGTGCTCGCG 235 10600 GCCAGCCGGGAGCTTCGGGT 236 10601 CCAGCCGGGAGCTTCGGGTG 237 10602 CAGCCGGGAGCTTCGGGTGC 238 10603 AGCCGGGAGCTTCGGGTGCT 239 10604 GCCGGGAGCTTCGGGTGCTC 240 10605 CCGGGAGCTTCGGGTGCTCG 241 10606 CGGGAGCTTCGGGTGCTCGC 242 10607 GGGAGCTTCGGGTGCTCGCG 243 10608 GGAGCTTCGGGTGCTCGCGC 244 10609 GAGCTTCGGGTGCTCGCGCT 245 10610 AGCTTCGGGTGCTCGCGCTG 246 10611 GCTTCGGGTGCTCGCGCTGC 247 10612 CTTCGGGTGCTCGCGCTGCT 248 10613 TTCGGGTGCTCGCGCTGCTT 249 10614 TCGGGTGCTCGCGCTGCTTC 250 10615 CGGGTGCTCGCGCTGCTTCT 251 10616 GGGTGCTCGCGCTGCTTCTC 252 10617 GGTGCTCGCGCTGCTTCTCC 253 10618 GTGCTCGCGCTGCTTCTCCG 254 10619 TGCTCGCGCTGCTTCTCCGG 255 10620 GCTCGCGCTGCTTCTCCGGG 256 10621 CTCGCGCTGCTTCTCCGGGT 257 10622 TCGCGCTGCTTCTCCGGGTA 258 10623 CGCGCTGCTTCTCCGGGTAC 259 10624 GCGCTGCTTCTCCGGGTACG 260 10625 CGCTGCTTCTCCGGGTACGG 261 10626 GCTGCTTCTCCGGGTACGGG 262 10627 CTGCTTCTCCGGGTACGGGC 263 10628 TGCTTCTCCGGGTACGGGCC 264 10629 GCTTCTCCGGGTACGGGCCG 265 10630 CTTCTCCGGGTACGGGCCGC 266 10631 TTCTCCGGGTACGGGCCGCT 267 10632 TCTCCGGGTACGGGCCGCTG 268 10633 CTCCGGGTACGGGCCGCTGG 269 10634 TCCGGGTACGGGCCGCTGGG 270 10635 CCGGGTACGGGCCGCTGGGT 271 10636 CGGGTACGGGCCGCTGGGTG 272 10637 GGGTACGGGCCGCTGGGTGG 273 10638 GGTACGGGCCGCTGGGTGGG 274 10639 GTACGGGCCGCTGGGTGGGG 275 10640 TACGGGCCGCTGGGTGGGGT 276 10641 ACGGGCCGCTGGGTGGGGTC 277 10642 CGGGCCGCTGGGTGGGGTCC 278 10643 GGGCCGCTGGGTGGGGTCCC 279 10644 GGCCGCTGGGTGGGGTCCCG 280 10645 GCCGCTGGGTGGGGTCCCGG 281 10646 CCGCTGGGTGGGGTCCCGGG 282 10647 CGCTGGGTGGGGTCCCGGGC 283 10648 GCTGGGTGGGGTCCCGGGCG 284 10649 CTGGGTGGGGTCCCGGGCGT 285 10650 TGGGTGGGGTCCCGGGCGTG 286 10651 GGGTGGGGTCCCGGGCGTGG 287 10652 GGTGGGGTCCCGGGCGTGGT 288 10653 GTGGGGTCCCGGGCGTGGTG 289 10654 TGGGGTCCCGGGCGTGGTGC 290 10655 GGGGTCCCGGGCGTGGTGCG 291 10656 GGGTCCCGGGCGTGGTGCGG 292 10657 GGTCCCGGGCGTGGTGCGGA 293 10658 GTCCCGGGCGTGGTGCGGAG 294 10659 TCCCGGGCGTGGTGCGGAGG 295 10660 CCCGGGCGTGGTGCGGAGGC 296 10661 CCGGGCGTGGTGCGGAGGCG 297 10662 CGGGCGTGGTGCGGAGGCGC 298 10663 CCGCCAGCCGGGAGCTTCGG 234 10664 GCCGCCAGCCGGGAGCTTCG 233 10665 TGCCGCCAGCCGGGAGCTTC 232 10666 CTGCCGCCAGCCGGGAGCTT 231 10667 TCTGCCGCCAGCCGGGAGCT 230 10668 TTCTGCCGCCAGCCGGGAGC 229 10669 TTTCTGCCGCCAGCCGGGAG 228 10670 GTTTCTGCCGCCAGCCGGGA 227 10671 GGTTTCTGCCGCCAGCCGGG 226 10672 CGGTTTCTGCCGCCAGCCGG 225 10673 CCGGTTTCTGCCGCCAGCCG 224 10674 CCCGGTTTCTGCCGCCAGCC 223 10675 TCCCGGTTTCTGCCGCCAGC 222 10676 CTCCCGGTTTCTGCCGCCAG 221 10677 ACTCCCGGTTTCTGCCGCCA 220 10678 CACTCCCGGTTTCTGCCGCC 219 10679 CCACTCCCGGTTTCTGCCGC 218 10680 CCCACTCCCGGTTTCTGCCG 217 10681 CCCCACTCCCGGTTTCTGCC 216 10682 GCCCCACTCCCGGTTTCTGC 215 10683 GGCCCCACTCCCGGTTTCTG 214 10684 CGGCCCCACTCCCGGTTTCT 213 10685 CCGGCCCCACTCCCGGTTTC 212 10686 CCCGGCCCCACTCCCGGTTT 211 10687 GCCCGGCCCCACTCCCGGTT 210 10688 CGCCCGGCCCCACTCCCGGT 209 10689 TCGCCCGGCCCCACTCCCGG 208 10690 CTCGCCCGGCCCCACTCCCG 207 10691 ACTCGCCCGGCCCCACTCCC 206 10692 CACTCGCCCGGCCCCACTCC 205 10693 GCACTCGCCCGGCCCCACTC 204 10694 CGCACTCGCCCGGCCCCACT 203 10695 GCGCACTCGCCCGGCCCCAC 202 10696 CGCGCACTCGCCCGGCCCCA 201 10697 CCGCGCACTCGCCCGGCCCC 200 10698 GCCGCGCACTCGCCCGGCCC 199 10699 TGCCGCGCACTCGCCCGGCC 198 10700 ATGCCGCGCACTCGCCCGGC 197 10701 GATGCCGCGCACTCGCCCGG 196 10702 GGATGCCGCGCACTCGCCCG 195 10703 GGGATGCCGCGCACTCGCCC 194 10704 TGGGATGCCGCGCACTCGCC 193 10705 CTGGGATGCCGCGCACTCGC 192 10706 CCTGGGATGCCGCGCACTCG 191 10707 GCCTGGGATGCCGCGCACTC 190 10708 GGCCTGGGATGCCGCGCACT 189 10709 CGGCCTGGGATGCCGCGCAC 188 10710 CCGGCCTGGGATGCCGCGCA 187 10711 GCCGGCCTGGGATGCCGCGC 186 10712 GGCCGGCCTGGGATGCCGCG 185 10713 GGGCCGGCCTGGGATGCCGC 184 10714 CGGGCCGGCCTGGGATGCCG 183 10715 TCGGGCCGGCCTGGGATGCC 182 10716 TTCGGGCCGGCCTGGGATGC 181 10717 GTTCGGGCCGGCCTGGGATG 180 10718 CGTTCGGGCCGGCCTGGGAT 179 10719 GCGTTCGGGCCGGCCTGGGA 178 10720 AGCGTTCGGGCCGGCCTGGG 177 10721 GAGCGTTCGGGCCGGCCTGG 176 10722 GGAGCGTTCGGGCCGGCCTG 175 10723 CGGAGCGTTCGGGCCGGCCT 174 10724 GCGGAGCGTTCGGGCCGGCC 173 10725 GGCGGAGCGTTCGGGCCGGC 172 10726 GGGCGGAGCGTTCGGGCCGG 171 10727 CGGGCGGAGCGTTCGGGCCG 170 10728 GCGGGCGGAGCGTTCGGGCC 169 10729 CGCGGGCGGAGCGTTCGGGC 168 10730 CCGCGGGCGGAGCGTTCGGG 167 10731 ACCGCGGGCGGAGCGTTCGG 166 10732 CACCGCGGGCGGAGCGTTCG 165 10733 CCACCGCGGGCGGAGCGTTC 164 10734 CCCACCGCGGGCGGAGCGTT 163 10735 GCCCACCGCGGGCGGAGCGT 162 10736 GGCCCACCGCGGGCGGAGCG 161 10737 CGGCCCACCGCGGGCGGAGC 160 10738 TCGGCCCACCGCGGGCGGAG 159 10739 GTCGGCCCACCGCGGGCGGA 158 10740 AGTCGGCCCACCGCGGGCGG 157 10741 AAGTCGGCCCACCGCGGGCG 156 10742 GAAGTCGGCCCACCGCGGGC 155 10743 GGAAGTCGGCCCACCGCGGG 154 10744 GGGAAGTCGGCCCACCGCGG 153 10745 GGGGAAGTCGGCCCACCGCG 152 10746 AGGGGAAGTCGGCCCACCGC 151 10747 GAGGGGAAGTCGGCCCACCG 150 10748 GGAGGGGAAGTCGGCCCACC 149 10749 CGGGTACGGGCCGCTGGGTGGGGTCCCG 272 10750 GGGTACGGGCCGCTGGGTGG 273 10751 GGTACGGGCCGCTGGGTGGG 274 10752 GTACGGGCCGCTGGGTGGGG 275 10753 TACGGGCCGCTGGGTGGGGT 276 10754 ACGGGCCGCTGGGTGGGGTC 277 10755 CGGGCCGCTGGGTGGGGTCC 278 10756 GGGCCGCTGGGTGGGGTCCC 279 10757 GGCCGCTGGGTGGGGTCCCG 280 10758 GCCGCTGGGTGGGGTCCCGG 281 10759 CCGCTGGGTGGGGTCCCGGG 282 10760 CGCTGGGTGGGGTCCCGGGC 283 10761 GCTGGGTGGGGTCCCGGGCG 284 10762 CTGGGTGGGGTCCCGGGCGT 285 10763 TGGGTGGGGTCCCGGGCGTG 286 10764 GGGTGGGGTCCCGGGCGTGG 287 10765 GGTGGGGTCCCGGGCGTGGT 288 10766 GTGGGGTCCCGGGCGTGGTG 289 10767 TGGGGTCCCGGGCGTGGTGC 290 10768 GGGGTCCCGGGCGTGGTGCG 291 10769 GGGTCCCGGGCGTGGTGCGG 292 10770 GGTCCCGGGCGTGGTGCGGA 293 10771 GTCCCGGGCGTGGTGCGGAG 294 10772 TCCCGGGCGTGGTGCGGAGG 295 10773 CCCGGGCGTGGTGCGGAGGC 296 10774 CCGGGCGTGGTGCGGAGGCG 297 10775 CGGGCGTGGTGCGGAGGCGC 298 10776 CCGGGTACGGGCCGCTGGGT 271 10777 TCCGGGTACGGGCCGCTGGG 270 10778 CTCCGGGTACGGGCCGCTGG 269 10779 TCTCCGGGTACGGGCCGCTG 268 10780 TTCTCCGGGTACGGGCCGCT 267 10781 CTTCTCCGGGTACGGGCCGC 266 10782 GCTTCTCCGGGTACGGGCCG 265 10783 TGCTTCTCCGGGTACGGGCC 264 10784 CTGCTTCTCCGGGTACGGGC 263 10785 GCTGCTTCTCCGGGTACGGG 262 10786 CGCTGCTTCTCCGGGTACGG 261 10787 GCGCTGCTTCTCCGGGTACG 260 10788 CGCGCTGCTTCTCCGGGTAC 259 10789 TCGCGCTGCTTCTCCGGGTA 258 10790 CTCGCGCTGCTTCTCCGGGT 257 10791 GCTCGCGCTGCTTCTCCGGG 256 10792 TGCTCGCGCTGCTTCTCCGG 255 10793 GTGCTCGCGCTGCTTCTCCG 254 10794 GGTGCTCGCGCTGCTTCTCC 253 10795 GGGTGCTCGCGCTGCTTCTC 252 10796 CGGGTGCTCGCGCTGCTTCT 251 10797 TCGGGTGCTCGCGCTGCTTC 250 10798 TTCGGGTGCTCGCGCTGCTT 249 10799 CTTCGGGTGCTCGCGCTGCT 248 10800 GCTTCGGGTGCTCGCGCTGC 247 10801 AGCTTCGGGTGCTCGCGCTG 246 10802 GAGCTTCGGGTGCTCGCGCT 245 10803 GGAGCTTCGGGTGCTCGCGC 244 10804 GGGAGCTTCGGGTGCTCGCG 243 10805 CGGGAGCTTCGGGTGCTCGC 242 10806 CCGGGAGCTTCGGGTGCTCG 241 10807 GCCGGGAGCTTCGGGTGCTC 240 10808 AGCCGGGAGCTTCGGGTGCT 239 10809 CAGCCGGGAGCTTCGGGTGC 238 10810 CCAGCCGGGAGCTTCGGGTG 237 10811 GCCAGCCGGGAGCTTCGGGT 236 10812 CGCCAGCCGGGAGCTTCGGG 235 10813 CCGCCAGCCGGGAGCTTCGG 234 10814 GCCGCCAGCCGGGAGCTTCG 233 10815 TGCCGCCAGCCGGGAGCTTC 232 10816 CTGCCGCCAGCCGGGAGCTT 231 10817 TCTGCCGCCAGCCGGGAGCT 230 10818 TTCTGCCGCCAGCCGGGAGC 229 10819 TTTCTGCCGCCAGCCGGGAG 228 10820 GTTTCTGCCGCCAGCCGGGA 227 10821 GGTTTCTGCCGCCAGCCGGG 226 10822 CGGTTTCTGCCGCCAGCCGG 225 10823 CCGGTTTCTGCCGCCAGCCG 224 10824 CCCGGTTTCTGCCGCCAGCC 223 10825 TCCCGGTTTCTGCCGCCAGC 222 10826 CTCCCGGTTTCTGCCGCCAG 221 10827 ACTCCCGGTTTCTGCCGCCA 220 10828 CACTCCCGGTTTCTGCCGCC 219 10829 CCACTCCCGGTTTCTGCCGC 218 10830 CCCACTCCCGGTTTCTGCCG 217 10831 CCCCACTCCCGGTTTCTGCC 216 10832 GCCCCACTCCCGGTTTCTGC 215 10833 GGCCCCACTCCCGGTTTCTG 214 10834 CGGCCCCACTCCCGGTTTCT 213 10835 CCGGCCCCACTCCCGGTTTC 212 10836 CCCGGCCCCACTCCCGGTTT 211 10837 GCCCGGCCCCACTCCCGGTT 210 10838 CGCCCGGCCCCACTCCCGGT 209 10839 TCGCCCGGCCCCACTCCCGG 208 10840 CTCGCCCGGCCCCACTCCCG 207 10841 ACTCGCCCGGCCCCACTCCC 206 10842 CACTCGCCCGGCCCCACTCC 205 10843 GCACTCGCCCGGCCCCACTC 204 10844 CGCACTCGCCCGGCCCCACT 203 10845 GCGCACTCGCCCGGCCCCAC 202 10846 CGCGCACTCGCCCGGCCCCA 201 10847 CCGCGCACTCGCCCGGCCCC 200 10848 GCCGCGCACTCGCCCGGCCC 199 10849 TGCCGCGCACTCGCCCGGCC 198 10850 ATGCCGCGCACTCGCCCGGC 197 10851 GATGCCGCGCACTCGCCCGG 196 10852 GGATGCCGCGCACTCGCCCG 195 10853 GGGATGCCGCGCACTCGCCC 194 10854 TGGGATGCCGCGCACTCGCC 193 10855 CTGGGATGCCGCGCACTCGC 192 10856 CCTGGGATGCCGCGCACTCG 191 10857 GCCTGGGATGCCGCGCACTC 190 10858 GGCCTGGGATGCCGCGCACT 189 10859 CGGCCTGGGATGCCGCGCAC 188 10860 CCGGCCTGGGATGCCGCGCA 187 10861 GCCGGCCTGGGATGCCGCGC 186 10862 GGCCGGCCTGGGATGCCGCG 185 10863 GGGCCGGCCTGGGATGCCGC 184 10864 CGGGCCGGCCTGGGATGCCG 183 10865 TCGGGCCGGCCTGGGATGCC 182 10866 TTCGGGCCGGCCTGGGATGC 181 10867 GTTCGGGCCGGCCTGGGATG 180 10868 CGTTCGGGCCGGCCTGGGAT 179 10869 GCGTTCGGGCCGGCCTGGGA 178 10870 AGCGTTCGGGCCGGCCTGGG 177 10871 GAGCGTTCGGGCCGGCCTGG 176 10872 GGAGCGTTCGGGCCGGCCTG 175 10873 CGGAGCGTTCGGGCCGGCCT 174 10874 GCGGAGCGTTCGGGCCGGCC 173 10875 GGCGGAGCGTTCGGGCCGGC 172 10876 GGGCGGAGCGTTCGGGCCGG 171 10877 CGGGCGGAGCGTTCGGGCCG 170 10878 GCGGGCGGAGCGTTCGGGCC 169 10879 CGCGGGCGGAGCGTTCGGGC 168 10880 CCGCGGGCGGAGCGTTCGGG 167 10881 ACCGCGGGCGGAGCGTTCGG 166 10882 CACCGCGGGCGGAGCGTTCG 165 10883 CCACCGCGGGCGGAGCGTTC 164 10884 CCCACCGCGGGCGGAGCGTT 163 10885 GCCCACCGCGGGCGGAGCGT 162 10886 GGCCCACCGCGGGCGGAGCG 161 10887 CGGCCCACCGCGGGCGGAGC 160 10888 TCGGCCCACCGCGGGCGGAG 159 10889 GTCGGCCCACCGCGGGCGGA 158 10890 AGTCGGCCCACCGCGGGCGG 157 10891 AAGTCGGCCCACCGCGGGCG 156 10892 GAAGTCGGCCCACCGCGGGC 155 10893 GGAAGTCGGCCCACCGCGGG 154 10894 GGGAAGTCGGCCCACCGCGG 153 10895 GGGGAAGTCGGCCCACCGCG 152 10896 AGGGGAAGTCGGCCCACCGC 151 10897 GAGGGGAAGTCGGCCCACCG 150 10898 GGAGGGGAAGTCGGCCCACC 149 10899 GTGCGGAGGCGCAGGGCCGGGCTCCG 306 10900 CTACGCGCGGCTGCAGGGGGCGC 339 10901 TACGCGCGGCTGCAGGGGGC 340 10902 ACGCGCGGCTGCAGGGGGCG 341 10903 CGCGCGGCTGCAGGGGGCGC 342 10904 GCGCGGCTGCAGGGGGCGCT 343 10905 CGCGGCTGCAGGGGGCGCTG 344 10906 GCGGCTGCAGGGGGCGCTGG 345 10907 CGGCTGCAGGGGGCGCTGGG 346 10908 CCTACGCGCGGCTGCAGGGG 338 10909 GCCTACGCGCGGCTGCAGGG 337 10910 TGCCTACGCGCGGCTGCAGG 336 10911 CTGCCTACGCGCGGCTGCAG 335 10912 TCTGCCTACGCGCGGCTGCA 334 10913 CTCTGCCTACGCGCGGCTGC 333 10914 TCTCTGCCTACGCGCGGCTG 332 10915 GTCTCTGCCTACGCGCGGCT 331 10916 CGTCTCTGCCTACGCGCGGC 330 10917 CCGTCTCTGCCTACGCGCGG 329 10918 TCCGTCTCTGCCTACGCGCG 328 10919 CTCCGTCTCTGCCTACGCGC 327 10920 GCTCCGTCTCTGCCTACGCG 326 10921 GGCTCCGTCTCTGCCTACGC 325 10922 GGGCTCCGTCTCTGCCTACG 324 10923 CGGGCTCCGTCTCTGCCTAC 323 10924 CCGGGCTCCGTCTCTGCCTA 322 10925 GCCGGGCTCCGTCTCTGCCT 321 10926 GGCCGGGCTCCGTCTCTGCC 320 10927 GGGCCGGGCTCCGTCTCTGC 319 10928 AGGGCCGGGCTCCGTCTCTG 318 10929 CAGGGCCGGGCTCCGTCTCT 317 10930 GCAGGGCCGGGCTCCGTCTC 316 10931 CGCAGGGCCGGGCTCCGTCT 315 10932 GCGCAGGGCCGGGCTCCGTC 314 10933 GGCGCAGGGCCGGGCTCCGT 313 10934 AGGCGCAGGGCCGGGCTCCG 312 10935 GAGGCGCAGGGCCGGGCTCC 311 10936 GGAGGCGCAGGGCCGGGCTC 310 10937 CGGAGGCGCAGGGCCGGGCT 309 10938 CTGCGCAGGACTCGCGTCCTGGCCCG 375 10939 TGCGCAGGACTCGCGTCCTG 376 10940 GCGCAGGACTCGCGTCCTGG 377 10941 CGCAGGACTCGCGTCCTGGC 378 10942 GCAGGACTCGCGTCCTGGCC 379 10943 CAGGACTCGCGTCCTGGCCC 380 10944 AGGACTCGCGTCCTGGCCCG 381 10945 GGACTCGCGTCCTGGCCCGG 382 10946 GACTCGCGTCCTGGCCCGGG 383 10947 ACTCGCGTCCTGGCCCGGGC 384 10948 CTCGCGTCCTGGCCCGGGCC 385 10949 TCGCGTCCTGGCCCGGGCCT 386 10950 CGCGTCCTGGCCCGGGCCTC 387 10951 GCGTCCTGGCCCGGGCCTCC 388 10952 CGTCCTGGCCCGGGCCTCCC 389 10953 GTCCTGGCCCGGGCCTCCCA 390 10954 TCCTGGCCCGGGCCTCCCAG 391 10955 CCTGGCCCGGGCCTCCCAGC 392 10956 CTGGCCCGGGCCTCCCAGCC 393 10957 TGGCCCGGGCCTCCCAGCCC 394 10958 GGCCCGGGCCTCCCAGCCCG 395 10959 GCCCGGGCCTCCCAGCCCGC 396 10960 CCCGGGCCTCCCAGCCCGCA 397 10961 CCGGGCCTCCCAGCCCGCAG 398 10962 CGGGCCTCCCAGCCCGCAGA 399 10963 GGGCCTCCCAGCCCGCAGAG 400 10964 GGCCTCCCAGCCCGCAGAGC 401 10965 GCCTCCCAGCCCGCAGAGCG 402 10966 CCTCCCAGCCCGCAGAGCGC 403 10967 CTCCCAGCCCGCAGAGCGCG 404 10968 TCCCAGCCCGCAGAGCGCGG 405 10969 CCCAGCCCGCAGAGCGCGGG 406 10970 CCAGCCCGCAGAGCGCGGGA 407 10971 CAGCCCGCAGAGCGCGGGAT 408 10972 AGCCCGCAGAGCGCGGGATG 409 10973 GCCCGCAGAGCGCGGGATGG 410 10974 CCCGCAGAGCGCGGGATGGC 411 10975 CCGCAGAGCGCGGGATGGCT 412 10976 CGCAGAGCGCGGGATGGCTC 413 10977 GCAGAGCGCGGGATGGCTCT 414 10978 CAGAGCGCGGGATGGCTCTG 415 10979 AGAGCGCGGGATGGCTCTGG 416 10980 GAGCGCGGGATGGCTCTGGG 417 10981 AGCGCGGGATGGCTCTGGGC 418 10982 GCGCGGGATGGCTCTGGGCT 419 10983 CGCGGGATGGCTCTGGGCTC 420 10984 GCGGGATGGCTCTGGGCTCA 421 10985 CGGGATGGCTCTGGGCTCAG 422 10986 GCTGCGCAGGACTCGCGTCC 374 10987 GGCTGCGCAGGACTCGCGTC 373 10988 CGGCTGCGCAGGACTCGCGT 372 10989 TCGGCTGCGCAGGACTCGCG 371 10990 CTCGGCTGCGCAGGACTCGC 370 10991 CCTCGGCTGCGCAGGACTCG 369 10992 ACCTCGGCTGCGCAGGACTC 368 10993 AACCTCGGCTGCGCAGGACT 367 10994 GAACCTCGGCTGCGCAGGAC 366 10995 GGAACCTCGGCTGCGCAGGA 365 10996 GGGAACCTCGGCTGCGCAGG 364 10997 GGGGAACCTCGGCTGCGCAG 363 10998 TGGGGAACCTCGGCTGCGCA 362 10999 CTGGGGAACCTCGGCTGCGC 361 11000 GCTGGGGAACCTCGGCTGCG 360 11001 CGCTGGGGAACCTCGGCTGC 359 11002 GCGCTGGGGAACCTCGGCTG 358 11003 GGCGCTGGGGAACCTCGGCT 357 11004 GGGCGCTGGGGAACCTCGGC 356 11005 GGGGCGCTGGGGAACCTCGG 355 11006 GGGGGCGCTGGGGAACCTCG 354 11007 AGGGGGCGCTGGGGAACCTC 353 11008 CAGGGGGCGCTGGGGAACCT 352 11009 CCCGCAGAGCGCGGGATGGCTC 411 11010 CCGCAGAGCGCGGGATGGCT 412 11011 CGCAGAGCGCGGGATGGCTC 413 11012 GCAGAGCGCGGGATGGCTCT 414 11013 CAGAGCGCGGGATGGCTCTG 415 11014 AGAGCGCGGGATGGCTCTGG 416 11015 GAGCGCGGGATGGCTCTGGG 417 11016 AGCGCGGGATGGCTCTGGGC 418 11017 GCGCGGGATGGCTCTGGGCT 419 11018 CGCGGGATGGCTCTGGGCTC 420 11019 GCGGGATGGCTCTGGGCTCA 421 11020 CGGGATGGCTCTGGGCTCAG 422 11021 GCCCGCAGAGCGCGGGATGG 410 11022 AGCCCGCAGAGCGCGGGATG 409 11023 CAGCCCGCAGAGCGCGGGAT 408 11024 CCAGCCCGCAGAGCGCGGGA 407 11025 CCCAGCCCGCAGAGCGCGGG 406 11026 TCCCAGCCCGCAGAGCGCGG 405 11027 CTCCCAGCCCGCAGAGCGCG 404 11028 CCTCCCAGCCCGCAGAGCGC 403 11029 GCCTCCCAGCCCGCAGAGCG 402 11030 GGCCTCCCAGCCCGCAGAGC 401 11031 GGGCCTCCCAGCCCGCAGAG 400 11032 CGGGCCTCCCAGCCCGCAGA 399 11033 CCGGGCCTCCCAGCCCGCAG 398 11034 CCCGGGCCTCCCAGCCCGCA 397 11035 GCCCGGGCCTCCCAGCCCGC 396 11036 GGCCCGGGCCTCCCAGCCCG 395 11037 TGGCCCGGGCCTCCCAGCCC 394 11038 CTGGCCCGGGCCTCCCAGCC 393 11039 CCTGGCCCGGGCCTCCCAGC 392 11040 TCCTGGCCCGGGCCTCCCAG 391 11041 GTCCTGGCCCGGGCCTCCCA 390 11042 CGTCCTGGCCCGGGCCTCCC 389 11043 GCGTCCTGGCCCGGGCCTCC 388 11044 CGCGTCCTGGCCCGGGCCTC 387 11045 TCGCGTCCTGGCCCGGGCCT 386 11046 CTCGCGTCCTGGCCCGGGCC 385 11047 ACTCGCGTCCTGGCCCGGGC 384 11048 GACTCGCGTCCTGGCCCGGG 383 11049 GGACTCGCGTCCTGGCCCGG 382 11050 AGGACTCGCGTCCTGGCCCG 381 11051 CAGGACTCGCGTCCTGGCCC 380 11052 GCAGGACTCGCGTCCTGGCC 379 11053 CGCAGGACTCGCGTCCTGGC 378 11054 GCGCAGGACTCGCGTCCTGG 377 11055 TGCGCAGGACTCGCGTCCTG 376 11056 CTGCGCAGGACTCGCGTCCT 375 11057 GCTGCGCAGGACTCGCGTCC 374 11058 GGCTGCGCAGGACTCGCGTC 373 11059 CGGCTGCGCAGGACTCGCGT 372 11060 TCGGCTGCGCAGGACTCGCG 371 11061 CTCGGCTGCGCAGGACTCGC 370 11062 CCTCGGCTGCGCAGGACTCG 369 11063 ACCTCGGCTGCGCAGGACTC 368 11064 AACCTCGGCTGCGCAGGACT 367 11065 GAACCTCGGCTGCGCAGGAC 366 11066 GGAACCTCGGCTGCGCAGGA 365 11067 GGGAACCTCGGCTGCGCAGG 364 11068 GGGGAACCTCGGCTGCGCAG 363 11069 TGGGGAACCTCGGCTGCGCA 362 11070 CTGGGGAACCTCGGCTGCGC 361 11071 GCTGGGGAACCTCGGCTGCG 360 11072 CGCTGGGGAACCTCGGCTGC 359 11073 GCGCTGGGGAACCTCGGCTG 358 11074 GGCGCTGGGGAACCTCGGCT 357 11075 GGGCGCTGGGGAACCTCGGC 356 11076 GGGGCGCTGGGGAACCTCGG 355 11077 GGGGGCGCTGGGGAACCTCG 354 11078 AGGGGGCGCTGGGGAACCTC 353 11079 CAGGGGGCGCTGGGGAACCT 352 11080 CGGACCTGATGGGGCACGGGCTTCCCC 448 11081 GGACCTGATGGGGCACGGGC 449 11082 GACCTGATGGGGCACGGGCT 450 11083 ACCTGATGGGGCACGGGCTT 451 11084 CCTGATGGGGCACGGGCTTC 452 11085 CTGATGGGGCACGGGCTTCC 453 11086 TGATGGGGCACGGGCTTCCC 454 11087 GATGGGGCACGGGCTTCCCC 455 11088 ATGGGGCACGGGCTTCCCCT 456 11089 TGGGGCACGGGCTTCCCCTT 457 11090 GGGGCACGGGCTTCCCCTTT 458 11091 GGGCACGGGCTTCCCCTTTT 459 11092 GGCACGGGCTTCCCCTTTTA 460 11093 GCACGGGCTTCCCCTTTTAA 461 11094 CACGGGCTTCCCCTTTTAAC 462 11095 ACGGGCTTCCCCTTTTAACG 463 11096 CGGGCTTCCCCTTTTAACGG 464 11097 GGGCTTCCCCTTTTAACGGT 465 11098 GGCTTCCCCTTTTAACGGTG 466 11099 GCTTCCCCTTTTAACGGTGG 467 11100 CTTCCCCTTTTAACGGTGGT 468 11101 TTCCCCTTTTAACGGTGGTT 469 11102 TCCCCTTTTAACGGTGGTTG 470 11103 CCCCTTTTAACGGTGGTTGG 471 11104 CCCTTTTAACGGTGGTTGGG 472 11105 CCTTTTAACGGTGGTTGGGG 473 11106 CTTTTAACGGTGGTTGGGGC 474 11107 TTTTAACGGTGGTTGGGGCC 475 11108 TTTAACGGTGGTTGGGGCCT 476 11109 TTAACGGTGGTTGGGGCCTA 477 11110 TAACGGTGGTTGGGGCCTAG 478 11111 AACGGTGGTTGGGGCCTAGA 479 11112 ACGGTGGTTGGGGCCTAGAA 480 11113 CGGTGGTTGGGGCCTAGAAG 481 11114 GCGGACCTGATGGGGCACGG 447 11115 AGCGGACCTGATGGGGCACG 446 11116 GAGCGGACCTGATGGGGCAC 445 11117 CGGTGGTTGGGGCCTAGAAGCG 481 11118 ACGGTGGTTGGGGCCTAGAA 480 11119 AACGGTGGTTGGGGCCTAGA 479 11120 TAACGGTGGTTGGGGCCTAG 478 11121 TTAACGGTGGTTGGGGCCTA 477 11122 TTTAACGGTGGTTGGGGCCT 476 11123 TTTTAACGGTGGTTGGGGCC 475 11124 CTTTTAACGGTGGTTGGGGC 474 11125 CCTTTTAACGGTGGTTGGGG 473 11126 CCCTTTTAACGGTGGTTGGG 472 11127 CCCCTTTTAACGGTGGTTGG 471 11128 TCCCCTTTTAACGGTGGTTG 470 11129 TTCCCCTTTTAACGGTGGTT 469 11130 CTTCCCCTTTTAACGGTGGT 468 11131 GCTTCCCCTTTTAACGGTGG 467 11132 GGCTTCCCCTTTTAACGGTG 466 11133 GGGCTTCCCCTTTTAACGGT 465 11134 CGGGCTTCCCCTTTTAACGG 464 11135 ACGGGCTTCCCCTTTTAACG 463 11136 CACGGGCTTCCCCTTTTAAC 462 11137 GCACGGGCTTCCCCTTTTAA 461 11138 GGCACGGGCTTCCCCTTTTA 460 11139 GGGCACGGGCTTCCCCTTTT 459 11140 GGGGCACGGGCTTCCCCTTT 458 11141 TGGGGCACGGGCTTCCCCTT 457 11142 ATGGGGCACGGGCTTCCCCT 456 11143 GATGGGGCACGGGCTTCCCC 455 11144 TGATGGGGCACGGGCTTCCC 454 11145 CTGATGGGGCACGGGCTTCC 453 11146 CCTGATGGGGCACGGGCTTC 452 11147 ACCTGATGGGGCACGGGCTT 451 11148 GACCTGATGGGGCACGGGCT 450 11149 GGACCTGATGGGGCACGGGC 449 11150 CGGACCTGATGGGGCACGGG 448 11151 GCGGACCTGATGGGGCACGG 447 11152 AGCGGACCTGATGGGGCACG 446 11153 GAGCGGACCTGATGGGGCAC 445 11154 CGCGCCCCTCGCTGTGACCGCCCAGCCCG 524 11155 GCGCCCCTCGCTGTGACCGC 525 11156 CGCCCCTCGCTGTGACCGCC 526 11157 GCCCCTCGCTGTGACCGCCC 527 11158 CCCCTCGCTGTGACCGCCCA 528 11159 CCCTCGCTGTGACCGCCCAG 529 11160 CCTCGCTGTGACCGCCCAGC 530 11161 CTCGCTGTGACCGCCCAGCC 531 11162 TCGCTGTGACCGCCCAGCCC 532 11163 CGCTGTGACCGCCCAGCCCG 533 11164 GCTGTGACCGCCCAGCCCGG 534 11165 CTGTGACCGCCCAGCCCGGC 535 11166 TGTGACCGCCCAGCCCGGCG 536 11167 GTGACCGCCCAGCCCGGCGT 537 11168 TGACCGCCCAGCCCGGCGTG 538 11169 GACCGCCCAGCCCGGCGTGG 539 11170 ACCGCCCAGCCCGGCGTGGC 540 11171 CCGCCCAGCCCGGCGTGGCC 541 11172 CGCCCAGCCCGGCGTGGCCC 542 11173 GCCCAGCCCGGCGTGGCCCA 543 11174 CCCAGCCCGGCGTGGCCCAA 544 11175 CCAGCCCGGCGTGGCCCAAA 545 11176 CAGCCCGGCGTGGCCCAAAT 546 11177 AGCCCGGCGTGGCCCAAATG 547 11178 GCCCGGCGTGGCCCAAATGC 548 11179 CCCGGCGTGGCCCAAATGCC 549 11180 CCGGCGTGGCCCAAATGCCA 550 11181 CGGCGTGGCCCAAATGCCAG 551 11182 GGCGTGGCCCAAATGCCAGC 552 11183 GCGTGGCCCAAATGCCAGCC 553 11184 CGTGGCCCAAATGCCAGCCA 554 11185 GCGCGCCCCTCGCTGTGACC 523 11186 TGCGCGCCCCTCGCTGTGAC 522 11187 CTGCGCGCCCCTCGCTGTGA 521 11188 ACTGCGCGCCCCTCGCTGTG 520 11189 AACTGCGCGCCCCTCGCTGT 519 11190 AAACTGCGCGCCCCTCGCTG 518 11191 CAAACTGCGCGCCCCTCGCT 517 11192 CCAAACTGCGCGCCCCTCGC 516 11193 CCCAAACTGCGCGCCCCTCG 515 11194 CCCCAAACTGCGCGCCCCTC 514 11195 ACCCCAAACTGCGCGCCCCT 513 11196 GACCCCAAACTGCGCGCCCC 512 11197 TGACCCCAAACTGCGCGCCC 511 11198 GTGACCCCAAACTGCGCGCC 510 11199 TGTGACCCCAAACTGCGCGC 509 11200 GTGTGACCCCAAACTGCGCG 508 11201 TGTGTGACCCCAAACTGCGC 507 11202 CTGTGTGACCCCAAACTGCG 506 11203 CGGGGAGTGGGACTGCGGCGGGGAGCCG 580 11204 TCGGGGAGTGGGACTGCGGC 579 11205 CTCGGGGAGTGGGACTGCGG 578 11206 ACTCGGGGAGTGGGACTGCG 577 11207 AACTCGGGGAGTGGGACTGC 576 11208 ACTCGCCGAAGGCCCCTGGGGAAC 718 11209 CTCGCCGAAGGCCCCTGGGG 719 11210 TCGCCGAAGGCCCCTGGGGA 720 11211 CGCCGAAGGCCCCTGGGGAA 721 11212 GCCGAAGGCCCCTGGGGAAC 722 11213 CCGAAGGCCCCTGGGGAACA 723 11214 CGAAGGCCCCTGGGGAACAT 724 11215 GACTCGCCGAAGGCCCCTGG 717 11216 AGACTCGCCGAAGGCCCCTG 716 11217 AAGACTCGCCGAAGGCCCCT 715 11218 AAAGACTCGCCGAAGGCCCC 714 11219 AAAAGACTCGCCGAAGGCCC 713 11220 AAAAAGACTCGCCGAAGGCC 712 11221 CAAAAAGACTCGCCGAAGGC 711 11222 CGGGCTGCATGCGTGAGCAGG 840 11223 GGGCTGCATGCGTGAGCAGG 841 11224 GGCTGCATGCGTGAGCAGGC 842 11225 GCTGCATGCGTGAGCAGGCT 843 11226 CTGCATGCGTGAGCAGGCTA 844 11227 TGCATGCGTGAGCAGGCTAG 845 11228 GCATGCGTGAGCAGGCTAGC 846 11229 CATGCGTGAGCAGGCTAGCA 847 11230 ATGCGTGAGCAGGCTAGCAG 848 11231 TGCGTGAGCAGGCTAGCAGC 849 11232 GCGTGAGCAGGCTAGCAGCA 850 11233 CGTGAGCAGGCTAGCAGCAG 851 11234 CCGGGCTGCATGCGTGAGCA 839 11235 CCCGGGCTGCATGCGTGAGC 838 11236 GCCCGGGCTGCATGCGTGAG 837 11237 AGCCCGGGCTGCATGCGTGA 836 11238 CAGCCCGGGCTGCATGCGTG 835 11239 GCAGCCCGGGCTGCATGCGT 834 11240 TGCAGCCCGGGCTGCATGCG 833 11241 CTGCAGCCCGGGCTGCATGC 832 11242 TCTGCAGCCCGGGCTGCATG 831 11243 CTCTGCAGCCCGGGCTGCAT 830 11244 CCTCTGCAGCCCGGGCTGCA 829 11245 TCCTCTGCAGCCCGGGCTGC 828 11246 TTCCTCTGCAGCCCGGGCTG 827 11247 CTTCCTCTGCAGCCCGGGCT 826 11248 ACTTCCTCTGCAGCCCGGGC 825 11249 CACTTCCTCTGCAGCCCGGG 824 11250 ACACTTCCTCTGCAGCCCGG 823 11251 CACACTTCCTCTGCAGCCCG 822 11252 CGGCAGGCGGTTTAGGCTGTGGCTG 885 11253 GGCAGGCGGTTTAGGCTGTG 886 11254 GCAGGCGGTTTAGGCTGTGG 887 11255 CAGGCGGTTTAGGCTGTGGC 888 11256 AGGCGGTTTAGGCTGTGGCT 889 11257 GGCGGTTTAGGCTGTGGCTG 890 11258 GCGGTTTAGGCTGTGGCTGA 891 11259 CGGTTTAGGCTGTGGCTGAC 892 11260 ACGGCAGGCGGTTTAGGCTG 884 11261 AACGGCAGGCGGTTTAGGCT 883 11262 GAACGGCAGGCGGTTTAGGC 882 11263 TGAACGGCAGGCGGTTTAGG 881 11264 CTGAACGGCAGGCGGTTTAG 880 11265 GCTGAACGGCAGGCGGTTTA 879 11266 GGCTGAACGGCAGGCGGTTT 878 11267 AGGCTGAACGGCAGGCGGTT 877 11268 CAGGCTGAACGGCAGGCGGT 876 11269 TCAGGCTGAACGGCAGGCGG 875 11270 CTCAGGCTGAACGGCAGGCG 874 11271 TCTCAGGCTGAACGGCAGGC 873 11272 CTCTCAGGCTGAACGGCAGG 872 11273 CCTCTCAGGCTGAACGGCAG 871 11274 GCCTCTCAGGCTGAACGGCA 870 11275 AGCCTCTCAGGCTGAACGGC 869 11276 CAGCCTCTCAGGCTGAACGG 868 11277 GCAGCCTCTCAGGCTGAACG 867 11278 CCGATTCGGATTGCTCCAGCTGG 962 11279 CGATTCGGATTGCTCCAGCT 963 11280 GATTCGGATTGCTCCAGCTG 964 11281 ATTCGGATTGCTCCAGCTGG 965 11282 TTCGGATTGCTCCAGCTGGT 966 11283 TCGGATTGCTCCAGCTGGTA 967 11284 CGGATTGCTCCAGCTGGTAA 968 11285 ACCGATTCGGATTGCTCCAG 961 11286 AACCGATTCGGATTGCTCCA 960 11287 TAACCGATTCGGATTGCTCC 959 11288 TTAACCGATTCGGATTGCTC 958 11289 CGCACCCACTCAGTTGCCACGGG 1008 11290 GCACCCACTCAGTTGCCACG 1009 11291 CACCCACTCAGTTGCCACGG 1010 11292 ACCCACTCAGTTGCCACGGG 1011 11293 CCCACTCAGTTGCCACGGGA 1012 11294 CCACTCAGTTGCCACGGGAC 1013 11295 CACTCAGTTGCCACGGGACA 1014 11296 ACTCAGTTGCCACGGGACAC 1015 11297 CTCAGTTGCCACGGGACACA 1016 11298 TCAGTTGCCACGGGACACAC 1017 11299 CAGTTGCCACGGGACACACC 1018 11300 AGTTGCCACGGGACACACCT 1019 11301 GTTGCCACGGGACACACCTG 1020 11302 TTGCCACGGGACACACCTGC 1021 11303 TGCCACGGGACACACCTGCT 1022 11304 GCCACGGGACACACCTGCTT 1023 11305 CCACGGGACACACCTGCTTT 1024 11306 CACGGGACACACCTGCTTTT 1025 11307 ACGGGACACACCTGCTTTTA 1026 11308 CGGGACACACCTGCTTTTAG 1027 11309 ACGCACCCACTCAGTTGCCA 1007 11310 CACGCACCCACTCAGTTGCC 1006 11311 TCACGCACCCACTCAGTTGC 1005 11312 TTCACGCACCCACTCAGTTG 1004 11313 TTTCACGCACCCACTCAGTT 1003 11314 TTTTCACGCACCCACTCAGT 1002 11315 CTTTTCACGCACCCACTCAG 1001 11316 CCTTTTCACGCACCCACTCA 1000 11317 CCCTTTTCACGCACCCACTC 999 11318 CCCCTTTTCACGCACCCACT 998 11319 CCCCCTTTTCACGCACCCAC 997 11320 CCCCCCTTTTCACGCACCCA 996 11321 TCCCCCCTTTTCACGCACCC 995 11322 ATCCCCCCTTTTCACGCACC 994 11323 GATCCCCCCTTTTCACGCAC 993 11324 TGATCCCCCCTTTTCACGCA 992 11325 ATGATCCCCCCTTTTCACGC 991 11326 GATGATCCCCCCTTTTCACG 990 11327 CGGAGACCCACAACGCAACACACC 1099 11328 GGAGACCCACAACGCAACAC 1100 11329 GAGACCCACAACGCAACACA 1101 11330 AGACCCACAACGCAACACAC 1102 11331 GACCCACAACGCAACACACC 1103 11332 ACCCACAACGCAACACACCT 1104 11333 CCCACAACGCAACACACCTG 1105 11334 CCACAACGCAACACACCTGA 1106 11335 CACAACGCAACACACCTGAA 1107 11336 ACAACGCAACACACCTGAAC 1108 11337 CAACGCAACACACCTGAACT 1109 11338 AACGCAACACACCTGAACTG 1110 11339 ACGCAACACACCTGAACTGG 1111 11340 CGCAACACACCTGAACTGGG 1112 11341 CCGGAGACCCACAACGCAAC 1098 11342 GCCGGAGACCCACAACGCAA 1097 11343 TGCCGGAGACCCACAACGCA 1096 11344 GTGCCGGAGACCCACAACGC 1095 11345 TGTGCCGGAGACCCACAACG 1094 11346 ATGTGCCGGAGACCCACAAC 1093 11347 AATGTGCCGGAGACCCACAA 1092 11348 AAATGTGCCGGAGACCCACA 1091 11349 GAAATGTGCCGGAGACCCAC 1090 11350 TGAAATGTGCCGGAGACCCA 1089 11351 CTGAAATGTGCCGGAGACCC 1088 11352 TCTGAAATGTGCCGGAGACC 1087 11353 CTCTGAAATGTGCCGGAGAC 1086 11354 CCTCTGAAATGTGCCGGAGA 1085 11355 GCCTCTGAAATGTGCCGGAG 1084 11356 AGCCTCTGAAATGTGCCGGA 1083 11357 GAGCCTCTGAAATGTGCCGG 1082 11358 TGAGCCTCTGAAATGTGCCG 1081

Hot Zones (Relative upstream location to gene start site)  1-750 800-1200

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11983) CTTTCCTCCAAGGACTGAAACAGACAAGGATACCCCCTCTTACCACTGTT ATTCTACATAGTGCAGAAAGTCCTGGCCAGAGCTATCAGGCAAGAGGAAG AAAGGAAGGGTATCCAAACTGGAAAGGAAGAAGTGGTGAGAAAGTTTTAA TTTTATTTTTTTGCATGTAGTTATTAAGTTTTCCTAGCACCATTTATTAA AGAGACTGTTTTTTTCCATTGTATGTTCTTTACAGCTTTGTCACAGATTA GTTGGTTGTAAGTGCATGGATTTATATGTGGATTCTCTATTCTGCTCCAT TGGTCTATGTGTCTGTTTCTATGCCAATACTGTGCTGTTTTGGTTACTAT AGCTTTGTAGTAAATTTTGAAGTTAGGTAGTGTGATGCCTCCAGCTTTGT TCTTTTTGCTCAGGATTGCTTTGGCTATTCAGGGTCTTTTGTGGTTTCTT ATAAATTTTAGGAATTTTTCTGTTTCGTGAAGAATGTCATTTGTATTTTG ATAGGAATTGCATTGAATCTGTAAATTGCTTTAGGTAGTATTGTAATTTT AACAATATTAGTTCTTCCAGTCTATCACGGAGTATATTTCTGGGTTTTTG TGTCCTCTTCGATTTCTTTTATCAGAGTTTTATTTATAGTTTTTCCTTGC ATAGATATTTTACTTTTTTAGTTAAATTGATTCCTAGGTATTGTACATTT TTGTAGCTATTGTAAAAGGAACTGCTTTCTTGATTTTTATTCAGATTGTT CACTGTTGTCATATAAATGCTATTGATTTTTGTATGTTGATTTTGCATCC TGCAACTTTACTGAATCAGATCTAACAGCTTTTAGGTGGACTCTTTAGAT TTTTCTAGGTATAAGATCATGTAGTCTGCAAACAAACCTAATTTGACTTC TTCCTTTCCAATTTGGATACCCTTCCTTTCTTCCTCTTGCCTGATAGCTC TGGCCAGGACTTTCTGTACTACGTAGAATAACAGTGGTGAGAGGGGGTAT CCTTGTCTGTTTCAGCCCTTGGAGGAAAGGCCTTCATTTTTTCTGTGTTC AGTATGATGTTGACTGTAGGTTTGTCATATATGGCCTTCATTATTTTGAA GTATGTTCCTTCTATATCCATTTTGGTGAGAATTTTTATCATAAAGGAAT GTTGAATTTTATCAAATGCTATCTCAGCATCTATTGAAATAAGTATACCG TTTCTGTTTTTGATTCTGTTAATGACTTATATTTAGATGGTATTTAAAGA CATAGGAAATGGGTTAGATCTCCTAAAGAGGGAAGATGGAAAGAGAATGA AAGAGTTTCCAGAATATAGCCCTGGGGGTTTCCCGCACTTATGGCGGAGG GAAGGGAGCCATCAGAGGAAACTCAGAGTGGCCAGATAGAAAGGCAGGAA AAAGCTTAATAAGGTGGTATCATGAGAAGAGAGTCTTCCTAGAAGGAGAA ATGCTTACTCTTGATAAAAACTGAAAAATAGGGTGAGTCCACATTAGAAT TCTCGCTACTGGGGAGGACTTTCTACACAAGGTTAGAGATCACTTGTATT TTGTCTACTATAATTTTAGTGCATAATGCATTTTTTTGGCATATAGTGGG CACTTATATACATATTAATCTGATGACATTTAATAATTGAGTGCCCAAAT ATTTATGTCGAGTTGAGAGCTAGGGATAGGCATGAGTCTTTCTTAAACAT CCTTCTCTTCTCCTTCCTCCACCCCCTCACTTGCCTTCAGAAGCATTGAT CAGAGAGATAGCAGTATCTTCAGTTTTTTTTAAAGCAACATGAAACACAC TTTATTCCTGCTAACATTAGGAAAAGCGAGCTGTTTTCCAAGCCCTGGAG GAAGGAAATTCAGCTAACTAACGTGAGGTAATGTAGGGTGGCTATTTCTT GAAAGGTAGTGAATCATAACTATAACCATACTATGGAAAAAAGTCCTGCC TTACCAACCACTCCACTGACTGCTTGTCACCAAAACTACGCTATGAAACG AATTGTGTTGAGTGGCTTTCATTGTAAAAGATTTTGGTGAAGGGAGGGAA AGAAACTGGTAGGGGTTCAGATCAGAAGATCTGGCTTTGCCAGTTTCTGG AGGGTGTCAGAATGGCTTCAACATACCTACTTCCTTGGCCTCAACTGGAG GTTTTGTAGCTGTAAACAAGAAGGATTGCATAGTTCAGAATAACGACACT GTAAGCTCATTGTGGAACTGGGTTAAAATCAGCATGTAGATCTACTAAGA AAGAAACACACTCAGCACTACTACAGAAAGAAACAGCCATGGGCCCTGAT TGTTAGCTTTCTGGAAGCCATTTCATTTTTACAATAGATTTATCACACAC TGTATTGACTTTTTCCAGTATAGAGTAAGAGAAAGTTAATATTCCTCATG TTTTGTCTGTTGACAGACTGAAAATAATTGCATTGAGTTTGGCTAGAATA TCCTGTCATTCCATAAACATCTCAAACTCCACATGGCTAAAACTTAACCC ATCCATGCCCCCATCTGCATGCACACATACATGCATATAACTTCATTTCT CAGTGTTTTTTTCTCCATGAATGGTAGCACCATTCTCTTTCAAGAAAGAG AAATACTTCCCCTTGGGATTATCCTATCTCTACTTATTGCTGCAGGGGCT TCCAAAATTAGGTTTTCTGTGTTCAGTCTTGCATTCACACTTCTGAAACC CAGAGCTGACCGAGACAAATTCTTCAACTTCCTGTCAGTCCCAACATAGA TTTAAAATTCCTAACCTGGCTCATGAGGTTCACCATGTATTCCTGCTTCC CTAAGTCAGCCTCATATCACACCAACCTGTGTGTGGAGTGGCTAAATACT CTAGCCAGGCAAGCTCTCTCAGTTCTTCTACCTGGCTCCTCTGGAGCCCT CCTTATTGCTCATCCCATTCTTAGCCTGATTCAAGATTCCTGGTCCTTCA AATCTCTCTTTAAGTGTCCTTACCTGGATCTTTCTCTAGTTAGTACAAAT TTTTCTATCTACCATTGGAGCGAACATTTTTTGAAACTTTGTATCAGTCC TGCCTTACTCTTGGTGGAATCCTGTGGTCCTAGTCAAGTGCCTGCTCCAT GAATGTGCTGAATAAATGAATAAGCATTTTAATTGTGTATCTGTCATTAG TGTCAGATGTGTTATTTATTCCAGCATGGTTTTAGCACACAGACACACTC TTTGATGCAGACTTTTCTTTTCTTTTTACATATAGCAACAATAAAAAACT AGACTTTCATCTCCTGAAAATATCAGTCTAATAATCACCTATGGCTGTCT CTCTGGTTGCTGAAGGAAAAAAAAAAAAAAGGCAGGGCACACCTGGATTG CATTAGAATGAGACTCACTACCCAGTTCAGGTGTGTTGCGTTGTGGGTCT CCGGCACATTTCAGAGGCTCATTAGGACCCTGACCCCACACTGGGGTTTA CACCCCTAAAAGCAGGTGTGTCCCGTGGCAACTGAGTGGGTGCGTGAAAA GGGGGGATCATCAATTACCAGCTGGAGCAATCCGAATCGGTTAAAGTGAA TCAAGTCACAGTGCTTCCTTAACCCAACCTCTCTGTTGGGGTCAGCCACA GCCTAAACCGCCTGCCGTTCAGCCTGAGAGGCTGCTGCTAGCCTGCTCAC GCATGCAGCCCGGGCTGCAGAGGAAGTGTGGGGAGGAAGGAAGTGGGTAT AGAAGGGTGCTGAGATGTGGGTCTTGAAGAGAATAGCCATAACGTCTTTG TCACTAAAATGTTCCCCAGGGGCCTTCGGCGAGTCTTTTTGTTTGGTTTT TTGTTTTTAATCTGTGGCTCTTGATAATTTATCTAGTGGTTGCCTACACC TGAAAAACAAGACACAGTGTTTAACTATCAACGAAAGAACTGGACGGCTC CCCGCCGCAGTCCCACTCCCCGAGTTTGTGGCTGGCATTTGGGCCACGCC GGGCTGGGCGGTCACAGCGAGGGGCGCGCAGTTTGGGGTCACACAGCTCC GCTTCTAGGCCCCAACCACCGTTAAAAGGGGAAGCCCGTGCCCCATCAGG TCCGCTCTTGCTGAGCCCAGAGCCATCCCGCGCTCTGCGGGCTGGGAGGC CCGGGCCAGGACGCGAGTCCTGCGCAGCCGAGGTTCCCCAGCGCCCCCTG CAGCCGCGCGTAGGCAGAGACGGAGCCCGGCCCTGCGCCTCCGCACCACG CCCGGGACCCCACCCAGCGGCCCGTACCCGGAGAAGCAGCGCGAGCACCC GAAGCTCCCGGCTGGCGGCAGAAACCGGGAGTGGGGCCGGGCGAGTGCGC GGCATCCCAGGCCGGCCCGAACGCTCCGCCCGCGGTGGGCCGACTTCCCC TCCTCTTCCCTCTCTCCTTCCTTTAGCCCGCTGGCGCCGGACACGCTGCG CCTCATCTCTTGGGGCGTTCTTCCCCGTTGGCCAACCGTCGCATCCCGTG CAACTTTGGGGTAGTGGCCGTTTAGTGTTGAATGTTCCCCACCGAGAGCG C ATG

TTR

Transthyretin is a 55 kDa protein that exists as a quaternary structure consisting of four monomers binding as two homodimers to create two thyroxine binding sites per tetramer. The dimer-dimer interface comes apart during the process of tetramer dissociation. TTR misfolding and aggregation is known to be associated with amyloid diseases such as senile systemic amyloidosis, familial amyloid polyneuropathy (FAP) and familial amyloid cardiomyopathy (Foss et al. 2005 Biochemistry 44 (47): 15525-33; Zeldenrust SR and Benson Md. (2010). Protein misfolding diseases: current and emerging principles and therapies. New York: Wiley. Westermark et al., Proc. Natl. Acad. Sci. U.S.A. 87 (7): 2843-5. TTR is predominantly synthesized in the liver and choroid plexus for secretion into blood and CNS, respectively. FAP is characterized by pain, paresthesia, muscular weakness, autonomic dysfunction due to the systemic deposition of variants of the transthyretin protein. A common mutations include the replacement of valine by methionine at position 30 (TTR V30M) or valine by isoleucine (TTR V122L). The misfolding of dissociated monomers is believed to cause aggregation into a variety of structures including amyloid fibrils. Treatment of familial TTR amyloid disease has historically relied on liver transplantation as a crude form of gene therapy. Recent approaches include molecules to kinetically stabilize the TTR tetramer or blocking the synthesis of TTR monomers by siRNA and antisense therapeutics.

Protein: TTR Gene: TTR (Homo sapiens, chromosome 18, 29171730-29178987 [NCBI Reference Sequence NC_(—)000018.9]; start site location: 29171866; strand: positive)

Gene Identification GeneID 7276 HGNC 12405 HPRD 01447 MIM 176300

Targeted Sequences Relative upstream Sequence Design location to gene start ID No: ID Sequence (5′-3′) site 11359 CAACGCCCTGGCTCGAGTGCAGTGGCACG 775 11432 CTACTATCTCAGATACTCGGCCAACTCG 1749 11450 CACGCGTTTCAGCACTGCACCCTGTTG 2086

Target Shift Sequences Relative upstream Sequence location ID to gene No: Sequence (5′-3′) start site 11359 CAACGCCCTGGCTCGAGTGCAGTGGCACG 775 11360 AACGCCCTGGCTCGAGTGCA 776 11361 ACGCCCTGGCTCGAGTGCAG 777 11362 CGCCCTGGCTCGAGTGCAGT 778 11363 GCCCTGGCTCGAGTGCAGTG 779 11364 CCCTGGCTCGAGTGCAGTGG 780 11365 CCTGGCTCGAGTGCAGTGGC 781 11366 CTGGCTCGAGTGCAGTGGCA 782 11367 TGGCTCGAGTGCAGTGGCAC 783 11368 GGCTCGAGTGCAGTGGCACG 784 11369 GCTCGAGTGCAGTGGCACGA 785 11370 CTCGAGTGCAGTGGCACGAT 786 11371 TCGAGTGCAGTGGCACGATC 787 11372 CGAGTGCAGTGGCACGATCA 788 11373 GAGTGCAGTGGCACGATCAC 789 11374 AGTGCAGTGGCACGATCACA 790 11375 GTGCAGTGGCACGATCACAG 791 11376 TGCAGTGGCACGATCACAGC 792 11377 GCAGTGGCACGATCACAGCT 793 11378 CAGTGGCACGATCACAGCTC 794 11379 AGTGGCACGATCACAGCTCG 795 11380 GTGGCACGATCACAGCTCGC 796 11381 TGGCACGATCACAGCTCGCT 797 11382 GGCACGATCACAGCTCGCTG 798 11383 GCACGATCACAGCTCGCTGC 799 11384 CACGATCACAGCTCGCTGCA 800 11385 ACGATCACAGCTCGCTGCAG 801 11386 CGATCACAGCTCGCTGCAGC 802 11387 GATCACAGCTCGCTGCAGCC 803 11388 ATCACAGCTCGCTGCAGCCT 804 11389 TCACAGCTCGCTGCAGCCTT 805 11390 CACAGCTCGCTGCAGCCTTG 806 11391 ACAGCTCGCTGCAGCCTTGA 807 11392 CAGCTCGCTGCAGCCTTGAC 808 11393 AGCTCGCTGCAGCCTTGACC 809 11394 GCTCGCTGCAGCCTTGACCT 810 11395 CTCGCTGCAGCCTTGACCTC 811 11396 TCGCTGCAGCCTTGACCTCC 812 11397 CGCTGCAGCCTTGACCTCCC 813 11398 GCTGCAGCCTTGACCTCCCG 814 11399 CTGCAGCCTTGACCTCCCGG 815 11400 TGCAGCCTTGACCTCCCGGG 816 11401 GCAGCCTTGACCTCCCGGGC 817 11402 CAGCCTTGACCTCCCGGGCT 818 11403 AGCCTTGACCTCCCGGGCTC 819 11404 GCCTTGACCTCCCGGGCTCA 820 11405 CCTTGACCTCCCGGGCTCAG 821 11406 CTTGACCTCCCGGGCTCAGG 822 11407 TTGACCTCCCGGGCTCAGGT 823 11408 TGACCTCCCGGGCTCAGGTC 824 11409 GACCTCCCGGGCTCAGGTCA 825 11410 ACCTCCCGGGCTCAGGTCAT 826 11411 CCTCCCGGGCTCAGGTCATC 827 11412 CTCCCGGGCTCAGGTCATCC 828 11413 TCCCGGGCTCAGGTCATCCT 829 11414 CCCGGGCTCAGGTCATCCTC 830 11415 CCGGGCTCAGGTCATCCTCC 831 11416 CGGGCTCAGGTCATCCTCCC 832 11417 CCAACGCCCTGGCTCGAGTG 774 11418 TCCAACGCCCTGGCTCGAGT 773 11419 CTCCAACGCCCTGGCTCGAG 772 11420 ACTCCAACGCCCTGGCTCGA 771 11421 CACTCCAACGCCCTGGCTCG 770 11422 TCACTCCAACGCCCTGGCTC 769 11423 CTCACTCCAACGCCCTGGCT 768 11424 TCTCACTCCAACGCCCTGGC 767 11425 GTCTCACTCCAACGCCCTGG 766 11426 GGTCTCACTCCAACGCCCTG 765 11427 GGGTCTCACTCCAACGCCCT 764 11428 AGGGTCTCACTCCAACGCCC 763 11429 CAGGGTCTCACTCCAACGCC 762 11430 ACAGGGTCTCACTCCAACGC 761 11431 GACAGGGTCTCACTCCAACG 760 11432 CTACTATCTCAGATACTCGGCCAACTCG 1749 11433 TACTATCTCAGATACTCGGC 1750 11434 ACTATCTCAGATACTCGGCC 1751 11435 CTATCTCAGATACTCGGCCA 1752 11436 TATCTCAGATACTCGGCCAA 1753 11437 ATCTCAGATACTCGGCCAAC 1754 11438 TCTCAGATACTCGGCCAACT 1755 11439 CTCAGATACTCGGCCAACTC 1756 11440 TCAGATACTCGGCCAACTCG 1757 11441 CAGATACTCGGCCAACTCGT 1758 11442 AGATACTCGGCCAACTCGTT 1759 11443 GATACTCGGCCAACTCGTTT 1760 11444 ATACTCGGCCAACTCGTTTG 1761 11445 TACTCGGCCAACTCGTTTGT 1762 11446 ACTCGGCCAACTCGTTTGTA 1763 11447 CTCGGCCAACTCGTTTGTAA 1764 11448 TCGGCCAACTCGTTTGTAAA 1765 11449 CGGCCAACTCGTTTGTAAAA 1766 11450 CACGCGTTTCAGCACTGCACCCTGTTG 2086 11451 ACGCGTTTCAGCACTGCACC 2087 11452 CGCGTTTCAGCACTGCACCC 2088 11453 GCGTTTCAGCACTGCACCCT 2089 11454 CGTTTCAGCACTGCACCCTG 2090 11455 GCACGCGTTTCAGCACTGCA 2085 11456 TGCACGCGTTTCAGCACTGC 2084 11457 GTGCACGCGTTTCAGCACTG 2083 11458 TGTGCACGCGTTTCAGCACT 2082 11459 CTGTGCACGCGTTTCAGCAC 2081 11460 ACTGTGCACGCGTTTCAGCA 2080 11461 TACTGTGCACGCGTTTCAGC 2079 11462 CTACTGTGCACGCGTTTCAG 2078 11463 TCTACTGTGCACGCGTTTCA 2077 11464 ATCTACTGTGCACGCGTTTC 2076 11465 AATCTACTGTGCACGCGTTT 2075 11466 AAATCTACTGTGCACGCGTT 2074 11467 AAAATCTACTGTGCACGCGT 2073 11468 CAAAATCTACTGTGCACGCG 2072 11469 GCAAAATCTACTGTGCACGC 2071 11470 AGCAAAATCTACTGTGCACG 2070

Hot Zones (Relative upstream location to gene start site) 735-915 1185-1275 1725-1815 2085-2175

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11984) AACTGGGCAGGCCTCAGGAAACTTACAATCATGGTAGAAGGTGAAGGGGA AGCAAAGCACCTTCCTCACAAGGCGTCAGGAAGAAGTGCCAAGCAAAGGG GGAAAAGCCCCTTGTAAAACTACCAGAACCTGTGAGAACTCAATCACTAT CACAAGAACAGCATGAGGGAACCGCCCCTCGTGATTCAATTACCTCCACC TGGTCTCTCCCTTGACACATGGGGATTATGGGTGTTACAATTCAAGATGA GATTTGGGTGGGGACACAAAGCCTAACCATATCAAGGATCAAGTGGTGGG TTGAAACTAACAGGATGAGATATATCAGATACAAACACAGGGTCCCATAT TTGGGTTAAAATTCATAAATGATCAAAGCACAGGATGACAGATAATATAG GTCATTTTAGATTATTGTGGCCAACAGATCACAGTGGGTAGTGTTATGAC GAAGGGAGGGTCACAGTTACTACAGTTACAGATGGATTCTGGGTACAACA TTTGCACTAAAGTGCCTTTGCCAAGGGAGGCAACAGTCTCGACATCCTGT GGCCTGATCTACTTCAGGGACTGTGTCTTGTTCAGAGCATCACATTTGAA GAGAACTTTGACCAAGGGGAATATGCCAGAAAAGGAAGTTCGGGATGCTG AGGATCTTAGGAACTATGTCTAAACAAGATTCATTCACAGAAGTGGGAAT GTCTATTTGGCAAAAAGAAAATACTACTTACATGGCTGTTGGAAGACCAG CAATCACAAACTCAGTTTTTCAAAAGGCTGGGCAGAAACACAGATGAAAG AAACAGGCCATGTTTAAGAAAAGATAAAAGCTCACGCATGATATGCCACT AGAGAATCACCTAGCCTCAGTGTTGGCGGGGAGGCCTGGGGAGTCTTGAT GTCTGAGAGTGACATTCTGATGATCACTGTCATGTGTAAATGTTGGCCTA AAGCTGCCAATATTTTTGATTTAAGAGAAGCAAGAAATGCAAATTTTTAT GCAGCATGTCTCAATTTTTAATTTTGGCAACTATTACAAAATGTTTAAAG AGACTCTGTGCAGCCCAAATATAACATATCTATGGGCTGATGGCAGCCCA GCGTTGCCAGTTCACAGGGTCTACAAGAGATGATTCTTAGTTTCAACAGG GTGCAGTGCTGAAACGCGTGCACAGTAGATTTTGCTTCGGTTATGAAAGA ACTTCCAAATATTTATGATTCATAGCCAGAGAAAAGGCTCTCTATCCAGG TTCTGAACAATAGGAAATCATCAAGAGGATATTGGATGACAATATATGAA AGATGTTATTTGAGAAAGGATTCTCTCCTGAGGCATAGATGTTGAACCAA ATTCTATTAGTTATGCTTTTACAGCAAGATAGTGGTTTACAGCTTACAAA AGGCTTGTACATCCTCTCATATTAAAAGTTATTAGAACAGTCCTTTGAAG TAGAAAAGTAGGCATTTCTATTTTACAAACGAGTTGGCCGAGTATCTGAG ATAGTAGATAACTCATAGAAGGTCATCCGGGAAACGGGGCAGCAGAACTG GGATCGAATGACTCTGGTCATCCAACTCCAAATGCAAAAGTCTTTCTGCT GCTGCTTCCTAGTTAAACTCTAAGGGTCTAAGACTCCATTCCTAGTTATG GTCTCAACTACATTTGCTCATTGCTGTGAGGGGTCAACCCACCTCCCGGA GTCCTCTCCTGCACATTCTCATGTTCCTGAAAGGCTTTTCTGTCCCTTCC ACTACTCCCTGTAAGCTCCTGTGCTTCACAATTTCTTGTTGAATTTTTTC TAATCTGACTCTATCAGTTATGGGAATGTTCCCTCAATTCTTAGTGCTCC AAACCGGACTTGCTCTTGGCTTGTATTTGTCCAAAATATTTGTCTTCTCT ATGTTTTCTACATGTTTGTCTTATAAGGACAAAAACCTGCCTTAGTTTAT CCATGAACAAAGCCACGCATGCTAGTGGACACACACACACATGCGCGTGC GCGCGCACACACACACACACACACATACACACAGAGACTTTGTATGTGAG TAATGAATCATCAAATCATCATAATTTCTGGACTTGTATTAATAAGTCGG CCAGGAGGAAAAGAATCTGCTGTCAATCATGGCTTCTGGTTCTCACAGTC ATCTCTACTTTCTTCCAGCAAGTTTGGTTCTGTCAAAAACCAGCTGTCAG CCTTGTTCCTGCATGCCCAATGCAGAAGAGTCAGTAAAGAAGATTTGGTT CTCTGTATTTCAGGGGCATCAATGCCAGGTTGAAATATGCCATTCTGGCC CAGCTCAGTGGCTCACACGTGTAATCCCAGCACTTTGGAAGGCCAAAGCG GGTGGATTGCTTGAGCTCAGGAGTTCGAGACCAGCCTGGGCAAGAGGCTG AGGTGGGAGGATGACCTGAGCCCGGGAGGTCAAGGCTGCAGCGAGCTGTG ATCGTGCCACTGCACTCGAGCCAGGGCGTTGGAGTGAGACCCTGTCAAAA AAAAAAAAAAAAAGGAAGGAAAAAAGGAAGGAAGGAAGGGAGGGAGGGAA GATGCCATTCTTAGATTGAAGTGGACTTTATCTGGGCAGAACACACACAC ACATACACACATGCACACACACATTGTGGAGAAATTGCTGACTAAGCAAA GCTTCCAAATGACTTAGTTTGGCTAAAATGTAGGCTTTTAAAAATGTGAG CACTGCCAAGGGTTTTTCCTTGTTGACCCATGGATCCATCAAGTGCAAAC ATTTTCTAATGCACTATATTTAAGCCTGTGCAGCTAGATGTCATTCAACA TGAAATACATTATTACAACTTGCATCTGTCTAAAATCTTGCATCTAAAAT GAGAGACAAAAAATCTATAAAAATGGAAAACATGCATAGAAATATGTGAG GGAGGAAAAAATTACCCCCAAGAATGTTAGTGCACGCAGTCACACAGGGA GAAGACTATTTTTGTTTTGTTTTGATTGTTTTGTTTTGTTTTGGTTGTTT TGTTTTGGTGACCTAACTGGTCAAATGACCTATTAAGAATATTTCATAGA ACGAATGTTCCGATGCTCTAATCTCTCTAGACAAGGTTCATATTTGTATG GGTTACTTATTCTCTCTTTGTTGACTAAGTCAATAATCAGAATCAGCAGG TTTGCAGTCAGATTGGCAGGGATAAGCAGCCTAGCTCAGGAGAAGTGAGT ATAAAAGCCCCAGGCTGGGAGCAGCCATCACAGAAGTCCACTCATTCTTG GCAGG ATG

CD68

CD68 (Cluster of Differentiation 68) is a glycoprotein that is expressed on monocytes/macrophages. It is often used as a marker for monocytes, histiocytes, giant cells, Kupffer cells, and osteoclasts. CD68 has been used to distinguish between diseases of similar appearance, e.g. (1) for monocytes of lymphoid origin and (2) macrophages to diagnose conditions related to proliferation or abnormality of these cells, such as malignant histiocytosis, histiocytic lymphoma, and Gaucher's disease. CD68 primarily localizes to lysosomes and endosomes with a smaller fraction circulating to the cell surface. It is a type I integral membrane protein with a heavily glycosylated extracellular domain and binds to tissue- and organ-specific lectins or selectins. The protein is also a member of the scavenger receptor family and has been reported to bind LDL. Scavenger receptors typically function to clear cellular debris, promote phagocytosis, and mediate the recruitment and -activation of macrophages. Alternative splicing of the gene results in multiple transcripts encoding different isoforms of CD68.

Protein: CD68 Gene: CD68 (Homo sapiens, chromosome 17, 7482805-7485429 [NCBI Reference Sequence: NC_(—)000017.10]; start site location: 7482996; strand: positive)

Gene Identification GeneID 968 HGNC 1693 MIM 153634

Targeted Sequences Relative upstream Sequence location to ID Sequence (5′-3′) gene start site 11989 CGAGAACATGGCTTTCCAGCGTCTG 520

Hot Zones (Relative upstream location to gene start site) 1-600

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11985) GCCACATTTGCCATATCGATTCTGCAGCAGATTGAATTAGATCTAAAAGC CACCCAGGCCTTGGTCCTAGCACCCACTCGAGAATTGGCTCAGCAGGTAA GAGTGGCTTCTATTCCCTCCTTCAGGGCTGATTTAGGGATGATGAGTATA ATCCAAGGACCAGAGAAGTCTTCTCTGATCACCACCTTGGGAGGAAGACA TGGGTGCCCTAACACTCTCGAGACCTGCTGGGTTAATTAAAAGCTATTTC TTACCCAAACGTAACCATTGCTTCCTCCACCCATTTCCTGAGTCAAATGG GAAAGCTGTTGGGTGAAGCCTGGCTGGCTGGGCAAGTTTGACTGTGTTCT GAATAAGCACCTTCACTATGGGCTAAGAGATCCCTTGGTGTGGGGGTGAT CTTACAGTAGTCAGAGCAGATGGACAGTCCTTTTCACCCTTGCTTAATAG CCAGAGCTGTTTCATGCCTGGGGCACACACAATTCTAATGCTGGACTTTT TCCTGGGTCATGCTGCAACACTGATGTCAGAGCATGTTTTTAAATGTTCT GTGGCAGGGGCAGTGATTATTCTGGGTGTGGATAATGTAAGAAGTTACAG CAGAGCTCCATTCTAAGGCACTTGGCTCTCAGTTTTCTCAGAGTGAACAT GCCTCGTAGCTTGGGTCCTATGGCAGGAGTGCAATAGGACATGGATATGC ATCACCTGTTCTATAAAACTGGTTGCTGGCTGGGTGTGGTGGCTCAACTC GTATAATCCCAACACTTTGGGAGGCCAAGGCAGGCAGATCTCTTGAGATC AGGAGTTGGAGACCAGCCTGGCCAACATAGTGAAACCCCGCTTCTACTAA AAATACAAAAATTAGCCAGGCATGGTGGCGTGTGCCTTTTATCCCAGCTA CTCGGGAAGCTCAGGCAGGAGAATTTAACCCAGGAGGTGGAGGTTGCAGT GAGCTGAGATTGTGCCATTGCACTCCAGCCTGGGCAACGAGCAAAGCTCT GTCTCAAAAAAAGAAAAAAAAAATGGTTGCTGCGTGATGAGGCAGTTGGT CAAATTAGTTTTCAGAAGGTTAAGGGTTCTAAATATCTAGAGTAAAGAAA CTGAATTAATTATCTGAGCGGCCTCATTGTGAATCACTGTACACTCAGGA ACCAGACTGAGTTGAAATCCTGTCTTTGCCACCTATTGACAGCACGATCT TAAGTGGATTTTAGCCTCTGCCTGTTTCTCAGCTGAATGTGAGTTTAATA ATAGTGCATGCCCCAAAGTTGTTGGTTAGGAATCAATACATGAAAAACAT TTAAGAATGGTGCCGGGCACAGTGGTAACTGACATATGAGCACCTGCCTC TCTCTGCTCAGATACAGAAGGTGGTCATGGCACTAGGAGACTACATGGGC GCCTCCTGTCACGCCTGTATCGGGGGCACCAACGTGCGTGCTGAGGTGCA GAAACTGCAGATGGAAGCTCCCCACATCATCGTGGGTACCCCTGGCCGTG TGTTTGATATGCTTAACCGGAGATACCTGTGTGAGTAATTCGGTTCTCCA ATCCCCTGGGTCACTTTGCTCTTGTGCACGCTTTCCAGTCTTTCAGCGTA AGCCAGAGTCATTCCCAAGGATGCTGGTTTCTCTCTGGGGGAAGAGCTGC TCTGTGATGGAGCCCATGCGTGTCATCTGAGCCTCTGGCTTCCCTGCCAG TGCAGCCCTGGCAGTGTCCTACTTCCCAGGGCTGTTGTCTGCCTGGCGGG AAGGTCCTGGGCAAAGGATCAGTCTTTGTACTCTGAGAGCAGACTACTTG GCTCCTCTCTGTTTTTTATCAGCGAAGTTGGATATATCTCTCCCACATTT CCCTAATCATATGCTATATATTGGCTTTTTTTTTCTTCTCTAGCCCCCAA ATACATCAAGATGTTTGTACTGGATGAAGCTGACGAAATGTTAAGCCGTG GATTCAAGGACCAGATCTATGACATATTCCAAAAGCTCAACAGCAACACC CAGGTGAGGGCAGTCTTGCTTGAATAGCTAATGATTCTTGAAAAATAGTA AGTGCCAGGGGAACCATATACTGGATTCTTGAGCCTTTTTATGCATCTGC TTCAGTTTTAGGTGTGGCTAGGGAAGGGAGCAGGCCTCAGGAAGGAACCA GCACTCTAAGACTGGCCTTTTTTTCCACTAGGTAGTTTTGCTGTCAGCCA CAATGCCTTCTGATGTGCTTGAGGTGACCAAGAAGTTCATGAGGGACCCC ATTCGGATTCTTGTCAAGAAGGAAGAGTTGACCCTGGAGGGTATCCGCCA GTTCTACATCAACGTGGAACGAGAGGTGGGGCCCAGTGCAGGAGGCGGGC CTGGTAGTGAGTTGTTGGGTATAGCCCCTGACTGATTTTTGTCCCCCAAC CTCCAGGAGTGGAAGCTGGACACACTATGTGACTTGTATGAAACCCTGAC CATCACCCAGGCAGTCATCTTCATCAACACCCGGAGGAAGGTGGACTGGC TCACCGAGAAGATGCATGCTCGAGATTTCACTGTATCCGCCATGGTGTGT TTGCCCGCTGCCAGCCTGTTGTGGGTCTGCCCGTCAGAAGTGTCCTACTT GAAGCCAGGGTTCCTGGAACCCAGGTGCCTACCTGGTCTGCTGCATATTT GTTTTCTCTTCCAGCATGGAGATATGGACCAAAAGGAACGAGACGTGATT ATGAGGGAGTTTCGTTCTGGCTCTAGCAGAGTTTTGATTACCACTGACCT GCTGGTGAGTAGAGGGAACTGATAGCAAAGGCAGAAGGGAGGATCCAAGG TGATTCCCTCTCCAAGGGGACATCAGTGCCTCTCAGGAAAGTAGCAGCTT GGAATAGAATCTGGCATGCCTAAGGCCTTTGGGGAACTGGGATGCTTATT TCCTCTGCCTTCCTTGGCTGCCCACATGGATGCCTAAGTGTCTTCCCTCC GGGATAGAGTGTCCTCCGTGCACATGCTGAAGAGTTGTCTTTCTTGACGT AGGCCAGAGGCATTGATGTGCAGCAGGTTTCTTTAGTCATCAACTATGAC CTTCCCACCAACAGGGAAAACTATATCCACAGGTAAGCGTAGATCTGGAA CACTCCCCTACCCCTTCACACCTGGCCCTCCCTGGGCTTAAAGCTCCTGA TATTCCTCATCCCCTTCCTTGTTTTCCAGAATCGGTCGAGGTGGACGGTT TGGCCGTAAAGGTGTGGCTATTAACATGGTGACAGAAGAAGACAAGAGGA CTCTTCGAGACATTGAGACCTTCTACAACACCTCCATTGAGGAAATGCCC CTCAATGTTGCTGACCTCATCTGAGGGGCTGTCCTGCCACCCAGCCCCAG CCAGGGCTCAATCTCTGGGGGCTGAGGAGCAGCAGGAGGGGGGAGGGAAG GGAGCCAAGGGATGGACATCTTGTCATTTTTTTTCTTTGAATAAATGTCA CTTTTTGAGGCAAAAGAAGGAACCGTGAACATTTTAGACACCCTTTTCTT TGGGGTAGGCTCTTGCCCCAGGCGCCGGCTCTTCTCCCAAAAAAAAAAAA AAAACACTAATCCATTTCCCTAACCTAGTAACCTCCAGATCCCAGAGGCT CTCCTCACCTCAGCTGAGCTCCTTTGAAAGTGATTCAAGGGACTATGTCA CTCAGCCTCATTTGCTGGACCAAATCTGGAGGGAGAACCCCTAAAACCCC TAAGTGAGGTTGCCCAGGGGGTTGTCCCCAGGTGGGGGGAAGCAGGGGAG AGAAAATGGTAGCCATTTTTACATTGTTTTGTATAGTATTTATTGATTCA GGAAACAAACACAAAATTCTGAATAAAATGACTTGGAAACTGCCTGTTTG GGCTTCTCATTTCTTACCTCCCCTTCCCTCTCCCACCTGCTACTGGGTGC ATCTCTGCTCCCCCCTTCCCCAGCAGATGGTTACCTTTGGGCTGTTGCTT TCTTGTCACCATCTGAGTTCTCAGACGCTGGAAAGCCATGTTCTCGGCTC TGTGAATGACAATGCTGACTGGAGTGCTGCCCCTCTGTAAAGGGCTGGGT GTGGATGGTCACAAGCCCCTCACATGCCTCAGCCAAGAGGAAGTAGTACA GGGGTCAGCCCAGAGGTCCAGGGGAAAGGAGTGGAAACCGATTTCCCCAC CAAGGGAGGGGCCTGTACCTCAGCTGTTCCCATAGCTACTTGCCACAACT GCCAAGCAAGTTTCGCTGAGTTTGACACATGGATCCCTGTGGATCAACTG CCCTAGGACTCCGTTTGCACCCATGTGACACTGTTGACTTTGCCCTGATG AAGCAGGGCCAACAGTCCCCTAACTTAATTACAAAAACTAATGACTAAGA GAGAGGTGGCTAGAGCTGAGGCCCCTGAGTCAGGCTGTGGGTGGGATCAT CTCCAGTACAGGAAGTGAGACTTTCATTTCCTCCTTTCCAAGAGAGGGCT GAGGGAGCAGGGTTGAGCAACTGGTGCAGACAGCCTAGCTGGACTTTGGG TGAGGCGGTTCAGCC ATG

ALK

Anaplastic lymphoma kinase (ALK) also known as ALK tyrosine kinase receptor or CD246 (cluster of differentiation 246) is an enzyme encoded by the ALK gene. ALK is believed to have a putative transmembrane domain and an extracellular domain. ALK is believed to have oncogenic properties in through several ways: mutations, amplified copies, or fusion products with other genes. The t(2; 5) chromosomal translocation is associated with approximately 60% anaplastic large-cell lymphomas (ALCLs) and creates a fusion gene consisting of the ALK gene and the nucleophosmin (NPM) gene: the 3′ half of ALK, derived from chromosome 2 and coding for the catalytic domain, is fused to the 5′ portion of NPM from chromosome 5. The product of NPM-ALK or EML4-ALK fusion genes are oncogenic in lymphoma and non-small cell lung cancers, respectively. In a smaller fraction of ALCL patients, the 3′ half of ALK is fused to the 5′ sequence of TPM3 gene, encoding for tropomyosin 3. In rare cases, ALK is fused to other 5′ fusion partners, such as TFG, ATIC, CLTC1, TPM4, MSN, ALO17, MYH9.

Protein: ALK Gene: ALK (Homo sapiens, chromosome 2, 29415640-30144477 [NCBI Reference Sequence: NC_(—)000002.11]; start site location: 30143525; strand: negative)

Gene Identification GeneID 238 HGNC 427 MIM 105590

Targeted Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 11471 CGCCGGAGGAGGCCGTTTACACTGC 3 11530 CGTGCGCGCAAGTCTCTTGCTTTCC 132 11555 CGCTCTCCGCGCCGAGTGCCGCGCC 269 11621 CGCCTTTTGCGTTCCTTTTGGCTCC 482 11681 CGCAGGCACTGGAGCGGCCCCGGCG 701 11794 CGACCCTCCGAACAGAGGCGGCGGG 851 11825 CGCGCTGCTGCCCGACCCACGCAGT 1022 11901 CGGGTCCGACTTCGGAAAAACAGGT 1313 11923 CGGCCTGTCGGGTAGCACAGGAGTT 2022

Targeted Shift Sequences Relative upstream Sequence location to ID No: Sequence (5′-3′) gene start site 11471 CGCCGGAGGAGGCCGTTTACACTGC 3 11472 GCCGGAGGAGGCCGTTTACA 4 11473 CCGGAGGAGGCCGTTTACAC 5 11474 CGGAGGAGGCCGTTTACACT 6 11475 GGAGGAGGCCGTTTACACTG 7 11476 GAGGAGGCCGTTTACACTGC 8 11477 AGGAGGCCGTTTACACTGCT 9 11478 GGAGGCCGTTTACACTGCTC 10 11479 GAGGCCGTTTACACTGCTCT 11 11480 AGGCCGTTTACACTGCTCTC 12 11481 GGCCGTTTACACTGCTCTCC 13 11482 GCCGTTTACACTGCTCTCCG 14 11483 CCGTTTACACTGCTCTCCGG 15 11484 CGTTTACACTGCTCTCCGGG 16 11485 GTTTACACTGCTCTCCGGGC 17 11486 TTTACACTGCTCTCCGGGCC 18 11487 TTACACTGCTCTCCGGGCCC 19 11488 TACACTGCTCTCCGGGCCCA 20 11489 ACACTGCTCTCCGGGCCCAG 21 11490 CACTGCTCTCCGGGCCCAGC 22 11491 ACTGCTCTCCGGGCCCAGCC 23 11492 CTGCTCTCCGGGCCCAGCCT 24 11493 TGCTCTCCGGGCCCAGCCTC 25 11494 GCTCTCCGGGCCCAGCCTCA 26 11495 CTCTCCGGGCCCAGCCTCAC 27 11496 TCTCCGGGCCCAGCCTCACC 28 11497 CTCCGGGCCCAGCCTCACCC 29 11498 TCCGGGCCCAGCCTCACCCT 30 11499 CCGGGCCCAGCCTCACCCTT 31 11500 CGGGCCCAGCCTCACCCTTC 32 11501 GGGCCCAGCCTCACCCTTCG 33 11502 GGCCCAGCCTCACCCTTCGC 34 11503 GCCCAGCCTCACCCTTCGCT 35 11504 CCCAGCCTCACCCTTCGCTC 36 11505 CCAGCCTCACCCTTCGCTCT 37 11506 CAGCCTCACCCTTCGCTCTC 38 11507 AGCCTCACCCTTCGCTCTCC 39 11508 GCCTCACCCTTCGCTCTCCC 40 11509 CCTCACCCTTCGCTCTCCCC 41 11510 CTCACCCTTCGCTCTCCCCG 42 11511 TCACCCTTCGCTCTCCCCGA 43 11512 CACCCTTCGCTCTCCCCGAG 44 11513 ACCCTTCGCTCTCCCCGAGA 45 11514 CCCTTCGCTCTCCCCGAGAT 46 11515 CCTTCGCTCTCCCCGAGATG 47 11516 CTTCGCTCTCCCCGAGATGG 48 11517 TTCGCTCTCCCCGAGATGGG 49 11518 TCGCTCTCCCCGAGATGGGA 50 11519 CGCTCTCCCCGAGATGGGAA 51 11520 GCTCTCCCCGAGATGGGAAG 52 11521 CTCTCCCCGAGATGGGAAGA 53 11522 TCTCCCCGAGATGGGAAGAG 54 11523 CTCCCCGAGATGGGAAGAGG 55 11524 TCCCCGAGATGGGAAGAGGC 56 11525 CCCCGAGATGGGAAGAGGCT 57 11526 CCCGAGATGGGAAGAGGCTC 58 11527 CCGAGATGGGAAGAGGCTCT 59 11528 CCGCCGGAGGAGGCCGTTTA 2 11529 CCCGCCGGAGGAGGCCGTTT 1 11530 CGTGCGCGCAAGTCTCTTGCTTTCC 132 11531 GTGCGCGCAAGTCTCTTGCT 133 11532 TGCGCGCAAGTCTCTTGCTT 134 11533 GCGCGCAAGTCTCTTGCTTT 135 11534 CGCGCAAGTCTCTTGCTTTC 136 11535 GCGCAAGTCTCTTGCTTTCC 137 11536 CGCAAGTCTCTTGCTTTCCC 138 11537 GCGTGCGCGCAAGTCTCTTG 131 11538 TGCGTGCGCGCAAGTCTCTT 130 11539 GTGCGTGCGCGCAAGTCTCT 129 11540 TGTGCGTGCGCGCAAGTCTC 128 11541 CTGTGCGTGCGCGCAAGTCT 127 11542 ACTGTGCGTGCGCGCAAGTC 126 11543 GACTGTGCGTGCGCGCAAGT 125 11544 GGACTGTGCGTGCGCGCAAG 124 11545 AGGACTGTGCGTGCGCGCAA 123 11546 GAGGACTGTGCGTGCGCGCA 122 11547 AGAGGACTGTGCGTGCGCGC 121 11548 CAGAGGACTGTGCGTGCGCG 120 11549 CCAGAGGACTGTGCGTGCGC 119 11550 TCCAGAGGACTGTGCGTGCG 118 11551 CTCCAGAGGACTGTGCGTGC 117 11552 TCTCCAGAGGACTGTGCGTG 116 11553 ATCTCCAGAGGACTGTGCGT 115 11554 GATCTCCAGAGGACTGTGCG 114 11555 CGCTCTCCGCGCCGAGTGCCGCGCC 269 11556 GCTCTCCGCGCCGAGTGCCG 270 11557 CTCTCCGCGCCGAGTGCCGC 271 11558 TCTCCGCGCCGAGTGCCGCG 272 11559 CTCCGCGCCGAGTGCCGCGC 273 11560 TCCGCGCCGAGTGCCGCGCC 274 11561 CCGCGCCGAGTGCCGCGCCC 275 11562 CGCGCCGAGTGCCGCGCCCC 276 11563 GCGCCGAGTGCCGCGCCCCC 277 11564 CGCCGAGTGCCGCGCCCCCG 278 11565 GCCGAGTGCCGCGCCCCCGT 279 11566 CCGAGTGCCGCGCCCCCGTC 280 11567 CGAGTGCCGCGCCCCCGTCT 281 11568 GAGTGCCGCGCCCCCGTCTG 282 11569 AGTGCCGCGCCCCCGTCTGT 283 11570 GTGCCGCGCCCCCGTCTGTA 284 11571 TGCCGCGCCCCCGTCTGTAG 285 11572 GCCGCGCCCCCGTCTGTAGC 286 11573 CCGCGCCCCCGTCTGTAGCT 287 11574 CGCGCCCCCGTCTGTAGCTC 288 11575 GCGCCCCCGTCTGTAGCTCG 289 11576 CGCCCCCGTCTGTAGCTCGC 290 11577 GCCCCCGTCTGTAGCTCGCT 291 11578 CCCCCGTCTGTAGCTCGCTG 292 11579 CCCCGTCTGTAGCTCGCTGC 293 11580 CCCGTCTGTAGCTCGCTGCG 294 11581 CCGTCTGTAGCTCGCTGCGC 295 11582 CGTCTGTAGCTCGCTGCGCT 296 11583 GTCTGTAGCTCGCTGCGCTC 297 11584 TCTGTAGCTCGCTGCGCTCG 298 11585 CTGTAGCTCGCTGCGCTCGG 299 11586 TGTAGCTCGCTGCGCTCGGT 300 11587 GTAGCTCGCTGCGCTCGGTA 301 11588 TAGCTCGCTGCGCTCGGTAC 302 11589 AGCTCGCTGCGCTCGGTACA 303 11590 GCTCGCTGCGCTCGGTACAG 304 11591 CTCGCTGCGCTCGGTACAGA 305 11592 TCGCTGCGCTCGGTACAGAG 306 11593 CGCTGCGCTCGGTACAGAGG 307 11594 GCTGCGCTCGGTACAGAGGA 308 11595 CTGCGCTCGGTACAGAGGAA 309 11596 TGCGCTCGGTACAGAGGAAC 310 11597 GCGCTCGGTACAGAGGAACT 311 11598 CGCTCGGTACAGAGGAACTA 312 11599 GCTCGGTACAGAGGAACTAC 313 11600 CTCGGTACAGAGGAACTACT 314 11601 TCGGTACAGAGGAACTACTA 315 11602 CGGTACAGAGGAACTACTAT 316 11603 CCGCTCTCCGCGCCGAGTGC 268 11604 CCCGCTCTCCGCGCCGAGTG 267 11605 TCCCGCTCTCCGCGCCGAGT 266 11606 CTCCCGCTCTCCGCGCCGAG 265 11607 CCTCCCGCTCTCCGCGCCGA 264 11608 GCCTCCCGCTCTCCGCGCCG 263 11609 AGCCTCCCGCTCTCCGCGCC 262 11610 GAGCCTCCCGCTCTCCGCGC 261 11611 TGAGCCTCCCGCTCTCCGCG 260 11612 TTGAGCCTCCCGCTCTCCGC 259 11613 CTTGAGCCTCCCGCTCTCCG 258 11614 CCTTGAGCCTCCCGCTCTCC 257 11615 ACCTTGAGCCTCCCGCTCTC 256 11616 GACCTTGAGCCTCCCGCTCT 255 11617 GGACCTTGAGCCTCCCGCTC 254 11618 GGGACCTTGAGCCTCCCGCT 253 11619 TGGGACCTTGAGCCTCCCGC 252 11620 CTGGGACCTTGAGCCTCCCG 251 11621 CGCCTTTTGCGTTCCTTTTGGCTCC 482 11622 GCCTTTTGCGTTCCTTTTGG 483 11623 CCTTTTGCGTTCCTTTTGGC 484 11624 CTTTTGCGTTCCTTTTGGCT 485 11625 TTTTGCGTTCCTTTTGGCTC 486 11626 TTTGCGTTCCTTTTGGCTCC 487 11627 TTGCGTTCCTTTTGGCTCCT 488 11628 TGCGTTCCTTTTGGCTCCTC 489 11629 GCGTTCCTTTTGGCTCCTCC 490 11630 CGTTCCTTTTGGCTCCTCCA 491 11631 CCGCCTTTTGCGTTCCTTTT 481 11632 GCCGCCTTTTGCGTTCCTTT 480 11633 GGCCGCCTTTTGCGTTCCTT 479 11634 TGGCCGCCTTTTGCGTTCCT 478 11635 CTGGCCGCCTTTTGCGTTCC 477 11636 CCTGGCCGCCTTTTGCGTTC 476 11637 TCCTGGCCGCCTTTTGCGTT 475 11638 GTCCTGGCCGCCTTTTGCGT 474 11639 TGTCCTGGCCGCCTTTTGCG 473 11640 CTGTCCTGGCCGCCTTTTGC 472 11641 GCTGTCCTGGCCGCCTTTTG 471 11642 CGCTGTCCTGGCCGCCTTTT 470 11643 ACGCTGTCCTGGCCGCCTTT 469 11644 CACGCTGTCCTGGCCGCCTT 468 11645 GCACGCTGTCCTGGCCGCCT 467 11646 TGCACGCTGTCCTGGCCGCC 466 11647 CTGCACGCTGTCCTGGCCGC 465 11648 GCTGCACGCTGTCCTGGCCG 464 11649 TGCTGCACGCTGTCCTGGCC 463 11650 CTGCTGCACGCTGTCCTGGC 462 11651 GCTGCTGCACGCTGTCCTGG 461 11652 AGCTGCTGCACGCTGTCCTG 460 11653 CAGCTGCTGCACGCTGTCCT 459 11654 CCAGCTGCTGCACGCTGTCC 458 11655 CCCAGCTGCTGCACGCTGTC 457 11656 TCCCAGCTGCTGCACGCTGT 456 11657 CTCCCAGCTGCTGCACGCTG 455 11658 GCTCCCAGCTGCTGCACGCT 454 11659 GGCTCCCAGCTGCTGCACGC 453 11660 CGGCTCCCAGCTGCTGCACG 452 11661 GCGGCTCCCAGCTGCTGCAC 451 11662 GGCGGCTCCCAGCTGCTGCA 450 11663 CGGCGGCTCCCAGCTGCTGC 449 11664 ACGGCGGCTCCCAGCTGCTG 448 11665 AACGGCGGCTCCCAGCTGCT 447 11666 GAACGGCGGCTCCCAGCTGC 446 11667 AGAACGGCGGCTCCCAGCTG 445 11668 GAGAACGGCGGCTCCCAGCT 444 11669 TGAGAACGGCGGCTCCCAGC 443 11670 CTGAGAACGGCGGCTCCCAG 442 11671 GCTGAGAACGGCGGCTCCCA 441 11672 GGCTGAGAACGGCGGCTCCC 440 11673 AGGCTGAGAACGGCGGCTCC 439 11674 AAGGCTGAGAACGGCGGCTC 438 11675 TAAGGCTGAGAACGGCGGCT 437 11676 TTAAGGCTGAGAACGGCGGC 436 11677 TTTAAGGCTGAGAACGGCGG 435 11678 TTTTAAGGCTGAGAACGGCG 434 11679 CTTTTAAGGCTGAGAACGGC 433 11680 ACTTTTAAGGCTGAGAACGG 432 11681 CGCAGGCACTGGAGCGGCCCCGGCG 701 11682 GCAGGCACTGGAGCGGCCCC 702 11683 CAGGCACTGGAGCGGCCCCG 703 11684 AGGCACTGGAGCGGCCCCGG 704 11685 GGCACTGGAGCGGCCCCGGC 705 11686 GCACTGGAGCGGCCCCGGCG 706 11687 CACTGGAGCGGCCCCGGCGG 707 11688 ACTGGAGCGGCCCCGGCGGC 708 11689 CTGGAGCGGCCCCGGCGGCA 709 11690 TGGAGCGGCCCCGGCGGCAG 710 11691 GGAGCGGCCCCGGCGGCAGC 711 11692 GAGCGGCCCCGGCGGCAGCA 712 11693 AGCGGCCCCGGCGGCAGCAG 713 11694 GCGGCCCCGGCGGCAGCAGC 714 11695 CGGCCCCGGCGGCAGCAGCT 715 11696 GGCCCCGGCGGCAGCAGCTG 716 11697 GCCCCGGCGGCAGCAGCTGA 717 11698 CCCCGGCGGCAGCAGCTGAG 718 11699 CCCGGCGGCAGCAGCTGAGG 719 11700 CCGGCGGCAGCAGCTGAGGG 720 11701 CGGCGGCAGCAGCTGAGGGC 721 11702 TCGCAGGCACTGGAGCGGCC 700 11703 TTCGCAGGCACTGGAGCGGC 699 11704 GTTCGCAGGCACTGGAGCGG 698 11705 AGTTCGCAGGCACTGGAGCG 697 11706 GAGTTCGCAGGCACTGGAGC 696 11707 AGAGTTCGCAGGCACTGGAG 695 11708 CAGAGTTCGCAGGCACTGGA 694 11709 TCAGAGTTCGCAGGCACTGG 693 11710 CTCAGAGTTCGCAGGCACTG 692 11711 CCTCAGAGTTCGCAGGCACT 691 11712 TCCTCAGAGTTCGCAGGCAC 690 11713 CTCCTCAGAGTTCGCAGGCA 689 11714 GCTCCTCAGAGTTCGCAGGC 688 11715 GGCTCCTCAGAGTTCGCAGG 687 11716 CGGCTCCTCAGAGTTCGCAG 686 11717 TCGGCTCCTCAGAGTTCGCA 685 11718 CTCGGCTCCTCAGAGTTCGC 684 11719 CCTCGGCTCCTCAGAGTTCG 683 11720 GCCTCGGCTCCTCAGAGTTC 682 11721 CGCCTCGGCTCCTCAGAGTT 681 11722 GCGCCTCGGCTCCTCAGAGT 680 11723 GGCGCCTCGGCTCCTCAGAG 679 11724 CGGCGCCTCGGCTCCTCAGA 678 11725 CCGGCGCCTCGGCTCCTCAG 677 11726 ACCGGCGCCTCGGCTCCTCA 676 11727 CACCGGCGCCTCGGCTCCTC 675 11728 TCACCGGCGCCTCGGCTCCT 674 11729 CTCACCGGCGCCTCGGCTCC 673 11730 TCTCACCGGCGCCTCGGCTC 672 11731 CTCTCACCGGCGCCTCGGCT 671 11732 GCTCTCACCGGCGCCTCGGC 670 11733 TGCTCTCACCGGCGCCTCGG 669 11734 TTGCTCTCACCGGCGCCTCG 668 11735 CTTGCTCTCACCGGCGCCTC 667 11736 CCTTGCTCTCACCGGCGCCT 666 11737 TCCTTGCTCTCACCGGCGCC 665 11738 GTCCTTGCTCTCACCGGCGC 664 11739 CGTCCTTGCTCTCACCGGCG 663 11740 GCGTCCTTGCTCTCACCGGC 662 11741 AGCGTCCTTGCTCTCACCGG 661 11742 CAGCGTCCTTGCTCTCACCG 660 11743 GCAGCGTCCTTGCTCTCACC 659 11744 TGCAGCGTCCTTGCTCTCAC 658 11745 TTGCAGCGTCCTTGCTCTCA 657 11746 TTTGCAGCGTCCTTGCTCTC 656 11747 GTTTGCAGCGTCCTTGCTCT 655 11748 AGTTTGCAGCGTCCTTGCTC 654 11749 AAGTTTGCAGCGTCCTTGCT 653 11750 CAAGTTTGCAGCGTCCTTGC 652 11751 GCAAGTTTGCAGCGTCCTTG 651 11752 CGCAAGTTTGCAGCGTCCTT 650 11753 GCGCAAGTTTGCAGCGTCCT 649 11754 TGCGCAAGTTTGCAGCGTCC 648 11755 CTGCGCAAGTTTGCAGCGTC 647 11756 GCTGCGCAAGTTTGCAGCGT 646 11757 CGCTGCGCAAGTTTGCAGCG 645 11758 GCGCTGCGCAAGTTTGCAGC 644 11759 CGCGCTGCGCAAGTTTGCAG 643 11760 CCGCGCTGCGCAAGTTTGCA 642 11761 CCCGCGCTGCGCAAGTTTGC 641 11762 CCCCGCGCTGCGCAAGTTTG 640 11763 CCCCCGCGCTGCGCAAGTTT 639 11764 GCCCCCGCGCTGCGCAAGTT 638 11765 AGCCCCCGCGCTGCGCAAGT 637 11766 CAGCCCCCGCGCTGCGCAAG 636 11767 CCAGCCCCCGCGCTGCGCAA 635 11768 CCCAGCCCCCGCGCTGCGCA 634 11769 TCCCAGCCCCCGCGCTGCGC 633 11770 ATCCCAGCCCCCGCGCTGCG 632 11771 AATCCCAGCCCCCGCGCTGC 631 11772 GAATCCCAGCCCCCGCGCTG 630 11773 TGAATCCCAGCCCCCGCGCT 629 11774 GTGAATCCCAGCCCCCGCGC 628 11775 CGTGAATCCCAGCCCCCGCG 627 11776 GCGTGAATCCCAGCCCCCGC 626 11777 GGCGTGAATCCCAGCCCCCG 625 11778 GGGCGTGAATCCCAGCCCCC 624 11779 TGGGCGTGAATCCCAGCCCC 623 11780 CTGGGCGTGAATCCCAGCCC 622 11781 TCTGGGCGTGAATCCCAGCC 621 11782 TTCTGGGCGTGAATCCCAGC 620 11783 CTTCTGGGCGTGAATCCCAG 619 11784 ACTTCTGGGCGTGAATCCCA 618 11785 AACTTCTGGGCGTGAATCCC 617 11786 GAACTTCTGGGCGTGAATCC 616 11787 TGAACTTCTGGGCGTGAATC 615 11788 CTGAACTTCTGGGCGTGAAT 614 11789 GCTGAACTTCTGGGCGTGAA 613 11790 TGCTGAACTTCTGGGCGTGA 612 11791 CTGCTGAACTTCTGGGCGTG 611 11792 CCTGCTGAACTTCTGGGCGT 610 11793 GCCTGCTGAACTTCTGGGCG 609 11794 CGACCCTCCGAACAGAGGCGGCGGG 851 11795 GACCCTCCGAACAGAGGCGG 852 11796 ACCCTCCGAACAGAGGCGGC 853 11797 CCCTCCGAACAGAGGCGGCG 854 11798 CCTCCGAACAGAGGCGGCGG 855 11799 CTCCGAACAGAGGCGGCGGG 856 11800 GCGACCCTCCGAACAGAGGC 850 11801 CGCGACCCTCCGAACAGAGG 849 11802 CCGCGACCCTCCGAACAGAG 848 11803 CCCGCGACCCTCCGAACAGA 847 11804 CCCCGCGACCCTCCGAACAG 846 11805 GCCCCGCGACCCTCCGAACA 845 11806 TGCCCCGCGACCCTCCGAAC 844 11807 GTGCCCCGCGACCCTCCGAA 843 11808 GGTGCCCCGCGACCCTCCGA 842 11809 CGGTGCCCCGCGACCCTCCG 841 11810 TCGGTGCCCCGCGACCCTCC 840 11811 CTCGGTGCCCCGCGACCCTC 839 11812 CCTCGGTGCCCCGCGACCCT 838 11813 ACCTCGGTGCCCCGCGACCC 837 11814 CACCTCGGTGCCCCGCGACC 836 11815 GCACCTCGGTGCCCCGCGAC 835 11816 AGCACCTCGGTGCCCCGCGA 834 11817 AAGCACCTCGGTGCCCCGCG 833 11818 AAAGCACCTCGGTGCCCCGC 832 11819 GAAAGCACCTCGGTGCCCCG 831 11820 GGAAAGCACCTCGGTGCCCC 830 11821 CGGAAAGCACCTCGGTGCCC 829 11822 CCGGAAAGCACCTCGGTGCC 828 11823 GCCGGAAAGCACCTCGGTGC 827 11824 GGCCGGAAAGCACCTCGGTG 826 11825 CGCGCTGCTGCCCGACCCACGCAGT 1022 11826 GCGCTGCTGCCCGACCCACG 1023 11827 CGCTGCTGCCCGACCCACGC 1024 11828 GCTGCTGCCCGACCCACGCA 1025 11829 CTGCTGCCCGACCCACGCAG 1026 11830 TGCTGCCCGACCCACGCAGT 1027 11831 GCTGCCCGACCCACGCAGTC 1028 11832 CTGCCCGACCCACGCAGTCC 1029 11833 TGCCCGACCCACGCAGTCCG 1030 11834 GCCCGACCCACGCAGTCCGG 1031 11835 CCCGACCCACGCAGTCCGGC 1032 11836 CCGACCCACGCAGTCCGGCC 1033 11837 CGACCCACGCAGTCCGGCCT 1034 11838 GACCCACGCAGTCCGGCCTC 1035 11839 ACCCACGCAGTCCGGCCTCG 1036 11840 CCCACGCAGTCCGGCCTCGC 1037 11841 CCACGCAGTCCGGCCTCGCC 1038 11842 CACGCAGTCCGGCCTCGCCC 1039 11843 ACGCAGTCCGGCCTCGCCCC 1040 11844 CGCAGTCCGGCCTCGCCCCG 1041 11845 GCAGTCCGGCCTCGCCCCGC 1042 11846 CAGTCCGGCCTCGCCCCGCC 1043 11847 AGTCCGGCCTCGCCCCGCCC 1044 11848 GTCCGGCCTCGCCCCGCCCC 1045 11849 TCCGGCCTCGCCCCGCCCCA 1046 11850 CCGGCCTCGCCCCGCCCCAC 1047 11851 CGGCCTCGCCCCGCCCCACC 1048 11852 GGCCTCGCCCCGCCCCACCC 1049 11853 GCCTCGCCCCGCCCCACCCG 1050 11854 CCTCGCCCCGCCCCACCCGC 1051 11855 CTCGCCCCGCCCCACCCGCA 1052 11856 TCGCCCCGCCCCACCCGCAC 1053 11857 CGCCCCGCCCCACCCGCACC 1054 11858 GCCCCGCCCCACCCGCACCC 1055 11859 CCCCGCCCCACCCGCACCCT 1056 11860 CCCGCCCCACCCGCACCCTC 1057 11861 CCGCCCCACCCGCACCCTCC 1058 11862 CGCCCCACCCGCACCCTCCA 1059 11863 GCCCCACCCGCACCCTCCAA 1060 11864 CCCCACCCGCACCCTCCAAC 1061 11865 CCCACCCGCACCCTCCAACC 1062 11866 CCACCCGCACCCTCCAACCA 1063 11867 CACCCGCACCCTCCAACCAA 1064 11868 ACCCGCACCCTCCAACCAAT 1065 11869 CCCGCACCCTCCAACCAATG 1066 11870 CCGCACCCTCCAACCAATGG 1067 11871 CGCACCCTCCAACCAATGGC 1068 11872 GCACCCTCCAACCAATGGCG 1069 11873 CACCCTCCAACCAATGGCGT 1070 11874 ACCCTCCAACCAATGGCGTG 1071 11875 CCCTCCAACCAATGGCGTGG 1072 11876 CCTCCAACCAATGGCGTGGC 1073 11877 CTCCAACCAATGGCGTGGCT 1074 11878 TCCAACCAATGGCGTGGCTC 1075 11879 CCAACCAATGGCGTGGCTCG 1076 11880 CAACCAATGGCGTGGCTCGA 1077 11881 AACCAATGGCGTGGCTCGAT 1078 11882 ACCAATGGCGTGGCTCGATC 1079 11883 CCGCGCTGCTGCCCGACCCA 1021 11884 TCCGCGCTGCTGCCCGACCC 1020 11885 CTCCGCGCTGCTGCCCGACC 1019 11886 ACTCCGCGCTGCTGCCCGAC 1018 11887 AACTCCGCGCTGCTGCCCGA 1017 11888 CAACTCCGCGCTGCTGCCCG 1016 11889 CCAACTCCGCGCTGCTGCCC 1015 11890 GCCAACTCCGCGCTGCTGCC 1014 11891 AGCCAACTCCGCGCTGCTGC 1013 11892 AAGCCAACTCCGCGCTGCTG 1012 11893 CAAGCCAACTCCGCGCTGCT 1011 11894 ACAAGCCAACTCCGCGCTGC 1010 11895 CACAAGCCAACTCCGCGCTG 1009 11896 TCACAAGCCAACTCCGCGCT 1008 11897 CTCACAAGCCAACTCCGCGC 1007 11898 GCTCACAAGCCAACTCCGCG 1006 11899 GGCTCACAAGCCAACTCCGC 1005 11900 GGGCTCACAAGCCAACTCCG 1004 11901 CGGGTCCGACTTCGGAAAAACAGGT 1313 11902 GGGTCCGACTTCGGAAAAAC 1314 11903 GGTCCGACTTCGGAAAAACA 1315 11904 GTCCGACTTCGGAAAAACAG 1316 11905 TCCGACTTCGGAAAAACAGG 1317 11906 CCGACTTCGGAAAAACAGGT 1318 11907 CGACTTCGGAAAAACAGGTT 1319 11908 GACTTCGGAAAAACAGGTTC 1320 11909 ACTTCGGAAAAACAGGTTCC 1321 11910 CTTCGGAAAAACAGGTTCCA 1322 11911 TTCGGAAAAACAGGTTCCAG 1323 11912 TCGGAAAAACAGGTTCCAGA 1324 11913 ACGGGTCCGACTTCGGAAAA 1312 11914 AACGGGTCCGACTTCGGAAA 1311 11915 AAACGGGTCCGACTTCGGAA 1310 11916 TAAACGGGTCCGACTTCGGA 1309 11917 TTAAACGGGTCCGACTTCGG 1308 11918 ATTAAACGGGTCCGACTTCG 1307 11919 GATTAAACGGGTCCGACTTC 1306 11920 AGATTAAACGGGTCCGACTT 1305 11921 GAGATTAAACGGGTCCGACT 1304 11922 AGAGATTAAACGGGTCCGAC 1303 11923 CGGCCTGTCGGGTAGCACAGGAGTT 2022 11924 GGCCTGTCGGGTAGCACAGG 2023 11925 GCCTGTCGGGTAGCACAGGA 2024 11926 CCTGTCGGGTAGCACAGGAG 2025 11927 CTGTCGGGTAGCACAGGAGT 2026 11928 TGTCGGGTAGCACAGGAGTT 2027 11929 GTCGGGTAGCACAGGAGTTT 2028 11930 TCGGGTAGCACAGGAGTTTT 2029 11931 CGGGTAGCACAGGAGTTTTC 2030 11932 ACGGCCTGTCGGGTAGCACA 2021 11933 CACGGCCTGTCGGGTAGCAC 2020 11934 TCACGGCCTGTCGGGTAGCA 2019 11935 CTCACGGCCTGTCGGGTAGC 2018 11936 GCTCACGGCCTGTCGGGTAG 2017 11937 AGCTCACGGCCTGTCGGGTA 2016 11938 GAGCTCACGGCCTGTCGGGT 2015 11939 GGAGCTCACGGCCTGTCGGG 2014 11940 TGGAGCTCACGGCCTGTCGG 2013 11941 CTGGAGCTCACGGCCTGTCG 2012 11942 TCTGGAGCTCACGGCCTGTC 2011 11943 CTCTGGAGCTCACGGCCTGT 2010 11944 TCTCTGGAGCTCACGGCCTG 2009 11945 CTCTCTGGAGCTCACGGCCT 2008 11946 CCTCTCTGGAGCTCACGGCC 2007 11947 TCCTCTCTGGAGCTCACGGC 2006 11948 ATCCTCTCTGGAGCTCACGG 2005 11949 GATCCTCTCTGGAGCTCACG 2004

Hot Zones (Relative upstream location to gene start site)  1-550 650-950 1000-1100 1250-1400 1950-2100

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 11986) TCTCTGCAGCCCCCTAGTGGCCATTGGGTGCAGCAGACGATTCACAGTTA ACTGACAAATTAACTGGAGTCAGTAATGCCTTTGGTCAAGAATTGTATAG AGAAATAGGGAAAGGCTGGAGTTTTAGTCTTTTTTCATATTTCAAATAAA AATTCCTCTTCCAGTAGGTATGTCAGAAAAATCTGATGAAAATCAAACAT ATATTGTACCAGGAAAGTATTAACTACCATAGCATTTTCCTCCCTCTTTT CTTTCTTTTCCACCCTTCCTCCACCAAGATAGGAGCATATTTTCTTCTCG GGTGAGATAATTCTTTGCCCTGAAACTTGTAAAGTCAGTGTATCCAGTGT GACTTCCAGAGAGAGGGCAGATGCCTGTCAAATTAAGTGAGTTGCCAAAC ATAGAGCAGGAAGAAAGCCATTCCGAGAATCAATATTCCTTTGTTACTGG GTCTTCCACTTGCCAAGGCATTGCCACAAAGCTGGAAAGGCCCAGCTCCT AGGAGAACAGAGGTTCCACCTGGCCACTATCTCCTGTGGGGTGGTAGGCA AGTTACTGCGGCCCCCAGGAGCTCAGTGAGGGAGGTTCAATGTGACACTG TGCTCTGATCCTGTGAGAAAACTCCTGTGCTACCCGACAGGCCGTGAGCT CCAGAGAGGATCTTGCCTTATTCTTAGCTTCAACAGTCAGCCCAAGGCCT GACAACCAGCCTTTAAGAAGGAATCAAGGGGATTTGTGTGACCCAAAGAT GGTAGTTTTGTCTGAGGATCTAGTGAACCACTTGTTATAAAAACAGCTAT TATGAGTTCTGTGTTGGCAGCTCAGGAGAGACGAAAGGAAAGGGAGAGGA GAGGTACAGCCATTACAGGTGAGTAAAAAAGGCCTAAGGTTCTGAACCCT CATTCCCAAGATTGTGGGCAAACAATTAAATGCTCTGCAACTCAGTTTCT GCATCTGTAAATCTGGAATTAAAATGTTTGCCTTACAGAGACTAGGGGAG GTTACACATGTTCAGACACCATTCTGAGAAAACAGAGCGACTGACAGGGG TCTGAAAGGTATTTGTTGTAGCTGCAGAACAACTCTGCCAGACCAAGACC ATCCATCCCTCTCTGCCCCCCTATTCCCAAATTCTCCTGTGTGGACGGCA GGACTCCTAAGCTCCCAGGAATGCATTCAAATAATAGATGGGTCAGAAAA TATTCTGTCTCAGGGCCTTAATACAAGCTGTTCTCAGATTTGCCAGTGTC GCGCTGCCACCCTCTCCCCACTTCCTCCTCCCTTCCCACTCCCCCCTCCC TTCCCCTCTCCTCCAGTTTTATTCTGGAACCTGTTTTTCCGAAGTCGGAC CCGTTTAATCTCTTAAATGTATAATTAGGGAGAGTGCTTGATTGCAAAGG CCTCTTCCAGTTCTCACATTTGCTCCCTTTCACACTGCAGAGAAATAGGG CAGGGAATCTAGAGGAGGGGAAGAACAAGAGACTGGAGAGGGAACAGAGG GAGGGTGGGGCGGGCTCACTCCTTTTCTCAATGAATGCCGAGGCCTCTGC AGATTTGCATAGGAGCCGATCGAGCCACGCCATTGGTTGGAGGGTGCGGG TGGGGCGGGGCGAGGCCGGACTGCGTGGGTCGGGCAGCAGCGCGGAGTTG GCTTGTGAGCCCCGCCCCCTCCGGGCCCCGCCCCCTCCCTGCGCGCGCTC GCGCGGCTCAGCCAGCTGCAAGTGGCGGGCGCCCAGGCAGATGCGATCCA GCGGCTCTGGGGGCGGCAGCGGTGGTAGCAGCTGGTACCTCCCGCCGCCT CTGTTCGGAGGGTCGCGGGGCACCGAGGTGCTTTCCGGCCGCCCTCTGGT CGGCCACCCAAAGCCGCGGGCGCTGATGATGGGTGAGGAGGGGGCGGCAA GATTTCGGGCGCCCCTGCCCTGAACGCCCTCAGCTGCTGCCGCCGGGGCC GCTCCAGTGCCTGCGAACTCTGAGGAGCCGAGGCGCCGGTGAGAGCAAGG ACGCTGCAAACTTGCGCAGCGCGGGGGCTGGGATTCACGCCCAGAAGTTC AGCAGGCAGACAGTCCGAAGCCTTCCCGCAGCGGAGAGATAGCTTGAGGG TGCGCAAGACGGCAGCCTCCGCCCTCGGTTCCCGCCCAGACCGGGCAGAA GAGCTTGGAGGAGCCAAAAGGAACGCAAAAGGCGGCCAGGACAGCGTGCA GCAGCTGGGAGCCGCCGTTCTCAGCCTTAAAAGTTGCAGAGATTGGAGGC TGCCCCGAGAGGGGACAGACCCCAGCTCCGACTGCGGGGGGCAGGAGAGG ACGGTACCCAACTGCCACCTCCCTTCAACCATAGTAGTTCCTCTGTACCG AGCGCAGCGAGCTACAGACGGGGGCGCGGCACTCGGCGCGGAGAGCGGGA GGCTCAAGGTCCCAGCCAGTGAGCCCAGTGTGCTTGAGTGTCTCTGGACT CGCCCCTGAGCTTCCAGGTCTGTTTCATTTAGACTCCTGCTCGCCTCCGT GCAGTTGGGGGAAAGCAAGAGACTTGCGCGCACGCACAGTCCTCTGGAGA TCAGGTGGAAGGAGCCGCTGGGTACCAAGGACTGTTCAGAGCCTCTTCCC ATCTCGGGGAGAGCGAAGGGTGAGGCTGGGCCCGGAGAGCAGTGTAAACG GCCTCCTCCGGCGGG A TG

Musashi Homolog 2 (MSI2)

Musashi homolog 2 is located on chromosome 17 and belongs to RNA-binding proteins of the Musashi family expressed in stem cell compartments and in aggressive tumors. MSI2 is the predominant form expressed in hematopoietic stem cells (HSCs), and its knockdown leads to reduced engraftment and depletion of HSCs in vivo. Overexpression of human MSI2 in a mouse model increases HSC cell cycle progression and cooperates with the chronic myeloid leukemia-associated BCR-ABL1 oncoprotein to induce an aggressive leukemia. MSI2 is overexpressed in human myeloid leukemia cell lines, and its depletion leads to decreased proliferation and increased apoptosis. Expression levels in human myeloid leukemia directly correlate with decreased survival in patients with the disease.

Protein: MSI2 Gene: MSI2 (Homo sapiens, chromosome 17, 57256570-57684689 [NCBI Reference Sequence: NC_(—)000017.11]; start site location: 57256743; strand: positive)

Gene Identification GeneID 124540 HGNC 18585 HPRD 07438 MIM 607897

Targeted Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 11989 CGGTGACGTCACGCACCCCCGTGCG 360 12058 CGGATACAATTACCCATATTGT 1535 12059 GACTCAGTTGCTAACAACCATGAGCG 10624 12060 CAGTTGCTAACAACCATGAGCG 10628 12061 CATGAAAATTTCACCAAGTATAAATTAC 10909 12062 CACCAAGTATAAATTACAGGTCT 10920

Targeted Shift Sequences Relative upstream location Sequence to gene ID No: Sequence (5′-3′) start site 11989 CGGTGACGTCACGCACCCCCGTGCG 354 11990 GGTGACGTCACGCACCCCCG 355 11991 GTGACGTCACGCACCCCCGT 356 11992 TGACGTCACGCACCCCCGTG 357 11993 GACGTCACGCACCCCCGTGC 358 11994 ACGTCACGCACCCCCGTGCG 359 11995 CGTCACGCACCCCCGTGCGG 360 11996 GTCACGCACCCCCGTGCGGC 361 11997 TCACGCACCCCCGTGCGGCC 362 11998 CACGCACCCCCGTGCGGCCC 363 11999 ACGCACCCCCGTGCGGCCCC 364 12000 CGCACCCCCGTGCGGCCCCC 365 12001 GCACCCCCGTGCGGCCCCCG 366 12002 CACCCCCGTGCGGCCCCCGC 367 12003 ACCCCCGTGCGGCCCCCGCC 368 12004 CCCCCGTGCGGCCCCCGCCT 369 12005 CCCCGTGCGGCCCCCGCCTG 370 12006 CCCGTGCGGCCCCCGCCTGC 371 12007 CCGTGCGGCCCCCGCCTGCC 372 12008 CGTGCGGCCCCCGCCTGCCC 373 12009 GTGCGGCCCCCGCCTGCCCG 374 12010 TGCGGCCCCCGCCTGCCCGC 375 12011 GCGGCCCCCGCCTGCCCGCG 376 12012 CGGCCCCCGCCTGCCCGCGC 377 12013 GGCCCCCGCCTGCCCGCGCG 378 12014 GCCCCCGCCTGCCCGCGCGC 379 12015 CCCCCGCCTGCCCGCGCGCG 380 12016 CCCCGCCTGCCCGCGCGCGC 381 12017 CCCGCCTGCCCGCGCGCGCA 382 12018 CCGCCTGCCCGCGCGCGCAC 383 12019 CGCCTGCCCGCGCGCGCACA 384 12020 GCCTGCCCGCGCGCGCACAC 385 12021 CCTGCCCGCGCGCGCACACT 386 12022 CTGCCCGCGCGCGCACACTC 387 12023 TGCCCGCGCGCGCACACTCG 388 12024 GCCCGCGCGCGCACACTCGG 389 12025 CCCGCGCGCGCACACTCGGC 390 12026 CCGCGCGCGCACACTCGGCC 391 12027 CGCGCGCGCACACTCGGCCC 392 12028 GCGCGCGCACACTCGGCCCC 393 12029 CGCGCGCACACTCGGCCCCC 394 12030 GCGCGCACACTCGGCCCCCC 395 12031 CGCGCACACTCGGCCCCCCA 396 12032 GCGCACACTCGGCCCCCCAC 397 12033 CGCACACTCGGCCCCCCACG 398 12034 GCACACTCGGCCCCCCACGG 399 12035 CACACTCGGCCCCCCACGGC 400 12036 ACACTCGGCCCCCCACGGCC 401 12037 CCGGTGACGTCACGCACCCC 353 12038 GCCGGTGACGTCACGCACCC 352 12039 TGCCGGTGACGTCACGCACC 351 12040 ATGCCGGTGACGTCACGCAC 350 12041 AATGCCGGTGACGTCACGCA 349 12042 CAATGCCGGTGACGTCACGC 348 12043 CCAATGCCGGTGACGTCACG 347 12044 ACCAATGCCGGTGACGTCAC 346 12045 AACCAATGCCGGTGACGTCA 345 12046 TAACCAATGCCGGTGACGTC 344 12047 GTAACCAATGCCGGTGACGT 343 12048 TGTAACCAATGCCGGTGACG 342 12049 GTGTAACCAATGCCGGTGAC 341 12050 CGTGTAACCAATGCCGGTGA 340 12051 TCGTGTAACCAATGCCGGTG 339 12052 GTCGTGTAACCAATGCCGGT 338 12053 CGTCGTGTAACCAATGCCGG 337 12054 ACGTCGTGTAACCAATGCCG 336 12055 AACGTCGTGTAACCAATGCC 335 12056 GAACGTCGTGTAACCAATGC 334 12057 AGAACGTCGTGTAACCAATG 333 12058 CGGATACAATTACCCATATTGT 1535 12059 GACTCAGTTGCTAACAACCATGAGCG 10624 12060 CAGTTGCTAACAACCATGAGCG 10628 12061 CATGAAAATTTCACCAAGTATAAATTAC 10909 12062 CACCAAGTATAAATTACAGGTCT 10920

Hot Zones (Relative upstream location to gene start site)  1-450 1450-1600 10000-11500

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 1364) ATTTCTCAAAGAACTAAAAATAGAACTGCCATTTGATCCAGCAATCCCAC TACTGGTAACCTTTAACAGTATATACCCAAAGGAAAAGAAATCAGTATAT CAAAAAGATACCCATACTCGTATGTTTATCGTAGCACTATTCACAATAGC AAAGATATGGAATCAACCTAAGTGTCCATCAACAGAGGATTGGATAAAGA AAATGTGATACATGTACACAATAAAGTACTACTCAGTCATTAAAAAAATC AAACAGCAGCAATATGGATGGAATTGCTGGAAGACATTATCCCCAGGTGA AACAAGCCGGAGACAGAAAGACAAACACTGCGTGTTTTCACTTATAAGTG GGAGCTAAATCATGTGTACACATGGATGTAGGGTGTGGAATAACAGATAA TGGAGACTTGAAAGAGTGAGGGGGCCAGGCATGGTGGCTCATGCCTGTAA TCCCAGCACTTTGGGAGGCCGAGGTGGGTGGATCATCTGAGGTCAGGAGT TTGAGACCAGCCTGGCCAACATGGTGAAACCTTGTCTCTACTAAAAATAC AAAAAGTAGCCGGGCATGGTGGTGTGTGCCTGTAATCCTAGCTACTCAGG AGGCTGAAGCAGGAGAATAGCTTGAACCCGGGAGGGTGGGAGGTTGCAGT GAGCCGAGATCACGCCACTGCACTCCAGCCTGGGCAACAAGAGCGAAACT CGGTCTCGAAAAAAGAAAAAAAAAAAAAAAAGAAAGGGTGAGGGGATGGG GGAAGTGAATGATGAGAGACTACTTAATGGGTACAATGTATTTGAGTGAT GAATACCCTAAAAACCCTGATTTTACCACTATGTGATCTATGCATGTAAC AAAATTATACACGTAACTCATAAATTTACATAAATAAACTAAAAAATAAT TTTTAAGTTAGCAAACAACTTTTTTAAAAGAAGAATAAGCAGATACCCCA CATAGTGAGTAGACAAAGAACATCCCAGGCAAAAGGAACAAACAGCATCT GCAAAGGGCTTGAAGAAGGAAACAGCTTGTTTTGTTTAAGAAATGATAGA AGGCTGGGCGTGGTGGCTCACGCCTGTAATCCTAGCACTTTGAGAGGCCA AGGAAGGTAGATTGCTTGAGCCTAAGAGTTTGGGACCAGCCTGGGCAACG TGGCAAAACCGCCAAAATTAACCGGGCTTGGTGGCATGCAGCTGTAGTCC CAGCTACTCAGGAGGCTGAGGTGGGAGGATCACCTGAGCCCAGGTGGTCA AGGCTGTGGTGTGCTGTGATCATGCTACTGCACTCCAGCCTGGATGACAG AGTGAGAACCTGTCTAAAAAATTAATTAATTAATTAAAAAAAATGAAAGA GATGATAGAAGATGGTGGTGTAATGTGAGGTTGGAAAAGCAGACCTAGAA CATACCACGGAAGGTCTTTTAGGTGACAGCAAGGGGGTTCGATGCAGTGG GAAACCGCTGGAGGGATCCGACCTGCATCCCATAAAGACCTCTTTGGCTA CTGTGAGGGAAACAGACATTTTGGGAAGGTTCCAGAAGTCAAGGTAGAGG AAGACTAGTCTATAAAGCGGACCGCCTTTGTGAAAAATCAACCTATGAAA GAAGCCAACATACAAAAGATACACTGGAAGTGATCAAAGACTTCCAAGAG CAGCCGGGAGGTGGAATCTCAAGTCCAGATGTTGAATGAGTTGGGTGATT GTATGGGACAGAACCAGAAGCTGATGAGGGGCCCAGGATGCAGATCAAAG AGATGGGATGAGCAGGCAAATGCCATTTCTTTTCATTCCGCCTATTTTGC TTGAGTCACCAGTTTGGTAAGGGGAGAGTTTCAACCATCTGCAAGTAATC CAAATGCTTTTACTAACCTGCCTACCCATCCACACCCCCACCAAAAAAAA AAAAAAAGAAAGAAAGAAAGAAAAATAAGAAGCCAACCCCAGAGCTTGTA CGCCTGCATTCTCCACAATCATTTCTCTGTGTTACAGCTCTTGTCATCTT ACATTATACATGAATAACTAATCAACCAAACACAAACCCACTGATGAAAA AAAACGCATGTTGGATCTAGAATGTGAGCAGGGTAAAGAGTAATATTAAT TTCACTGGCAAATAACAAAATGGGAGCAAACATGAGGGATGTTAAGACAA GACTTGTGTTTTTGAATGCTTCTGGGAATGGAAACTCTCCAAGAATCAAG GGAAAGGGGAAGAAGGTTGGACATATTTGGAGTGCACTAATACCAATTCT TTTTTCTTCTGGTATGCATATGTGACTATATGCAACATCATCCTGGGTCA GGCCAACTGGTCATTTAAACCACCATGCTGTCTGCCTCACCTAGTCGAAC AGTTCATATAATGATCTTCCACTATTCCTTAATGGACATACAGGTGTTAG TCTTAATGCACCAACCAATATTGCGATAGGCTGGAACAAAACCTTTTGCT CTTTGAATGTCCACAGTGAGTAACACTATGCTAGGTATGTAGTAGGTGCT CAATAAATGTATCTTTCACACACTTCCTAAATGAAGTCTATTTCTTCCTT CATCGTTTTCCTGAAAGATCTTTCTTGACCACTAAGGATGAACTCCTCAT CTCCATGTCCAGTGAGTGGTTTAGAAATGTTTTAGTCTTCCTCCTCCTTG ACTTCATAATGGCATCTGACACTGCTGTGACACTTCAGTTGTCTTCTGTG ACTCTGAACTTTTCTGGTTCTAATCTTTTCTCCCTGATGGCCTCTCTACC CTGACCTCCAATGATTTCTTTTCCTGTTTCTGCACTCGAAAGTCCATATT CCACAAGGACTCTGCTTTTCTGTCTTGTCACTCTTTATGAGAAATTAATT GTATCTCATTTCAAATCAATGCATGTATCTATCATCCTCTGTCACATTAC ATGCAAAGTATCTGCTTATGGATCTGTCTGTCCAACTAGACCTCAAGCTC CTCAAAAACAGATCCCTCACTGTGTTCGTCTCTGTGCCTCCAGTACCTGG CACAGAGCTGGACACTCAGTGAATGCTCCATACACACGGGCCCAACTGAT CCATTCTCAAATTATGCTCGGAGTTGGTAGATTGGGCAGAAATTATTACC CCCATTATATAGAGAGGGGACCCCAAAGCTCAAGGAGGTAAAGCAAACAT TAGAAACATGATTCCTGTGACTGGCTGTGGAGACTCCGTGTAATAATCAT TCTGGTATTGCCAATTGCAAAAGGACTGTAAAACTAATAGAGATGCTGTC TAAATACCGACATCCAATCCTTTCATGCTCTTCTGAGCAGGCTTGAATTT TCCAGTGCCTCTCTTAAAATACAGCCCGCAGGACTGAGCATAAAACTGCA TGTGGCCTGACCTGCCAGCACGCGCTGGGAGTATTGCTTCCCTCCTTCCA CACTCTGCTGTGCTATTAATAGAGCCTCAGATTGCACTAGGGTTCTCAAC AGCCTCCTCGCGCTGTAGGTTCACATCAAGCTGGTGATCAGCTAACCCCC CGGGTCTTTCTCATGCAATGCCTTCTGTCAGGCCAGGCCTCCCCAGCAGC CACTTGGGCAATGAATGTTTTTGAATGCTCATGATTATTTGTGACTTAGA CTAGAAGGTACAACACCAGATCATGCCCTTCTTTCTCATGTGGCATTTTG CTGAGTGGGTGATGGGGTCTTAGGGGTCCTGTTCAGGACCCAGAGCTGTG GCCAGCCACTGGGGGCCACCAGCATCATCGGCCCCACAAAAGTAAGAGGA TGATGGCGAAATTAATTTTCTGGCTCACTTCAAATTATTCTTATGTTCAT CCCTACTCCTCTTGGATTAGGTCCTTTCTTGGTCTTGTTCTAATGCTGGG TGTTTGTATGCACAGCCTCTTTTCCCTCCCACCCATGGACACAGCCACAT TTTGATTTCTCATGCCCTTTCCAAATAGTAGTAGTATTTTTAGCATGAAT ATCTTGCTCAGAAATTGGCTGTACAGTCTATCCCCTATTGTCTTCACATC TAATACTTTTCTTTTTTCCCTCTGCCTCTCCTATGCATCAATACCACGTT TGGCTAAAGAGATTTGATTTTGACCTATTTGAATTCTCCCTTCACAGATG ACAGCCCTTTCTCCCTCTCCTTCCTTCCCTTCAATTAGTTTTTTCAGCCA CTTGGAATTAGCTGATGGGTGATTGTAGAAATTGCAACGTGGGCTACTGT GGTGGGTCTCAATGTCAACTCCAGGAAACCTCTGAATCTGGGGGGCTCTG GTTTCAGAATCTGATAGCCAGGCCCTGAACTCTGGAATGTGGGGCTGTGA CATGAGATACAGCTTCTCTTTGCTCGGCCACTAGAGGGAACTGAAATGTG GGGCGCAAGAAGGCGTTTCCTCTGTTCGCCAGACGAGGGCGCTCATACCC ATGGTCCTCCAAAGTTGGAATTTCTCGCCCCAAAACAGATATTGTCGGGT TGGCCTCCTTGTAACCCAAACATACGATGGGATGACATTACAGCTGTGCT ACTGATTGCTGCTTTGACCGCCTCCTATGCTGTGTAAATGGCCAAAAGCA AAGAATTATTAAAAAGCAGGCCCAATGTTGTCCAAGCTCACGTGTGGTTT GTGGGTCTATGTGTTTGCTGCTGGCAAATTTGCAAGCAGATGGGACTCCA AGGCAAGGCGTGGAAGTGATGATGGGAACGTTGGAAGTTCACAGACATAA CTTGTAGAGTGTGTGAGGCCGGGTGCGCGGACCCTGTGTATCTGCAGCTG CGATACTTAGATTTCAGTTTGGCAAGGCAGGTCACGGTGGAGATGGGGCA AGCTGCAAGGGTGGTGGAGAGGAGGAAGGGAAGGTGACAGTGGCCCTCTG TCAACTGTTTCCAGGTGGAGTTGAAAGGTGTAATCATTTTCTTCTGGGGG CCTTGGCACCTTTCATCAAGACGAAGTTGGTGACTGGTTTAAAAAGATTT AAAAAATTAAGCTCGAGAGGCCAAAGGAGAAAATGGTTTCCAGGTGGAAA GGGCTTGACAGAATGGTGCTCTTGTGCCGTGACTCCGAACTCCGTGGAGC ATTCCAGTGGCCCACTGTACTCCCACCCCTCCAGGCAGCACTGGGAGGCA GCCAAGTCTAGGAGGCAAAGGGCTCCCTAACTGCCAAGCAGTGAAGATGT TGAATAAAATATTTACTTACACGTTTAAGAATAATGATGACAGCATGACA AACAGTGGTGAAACAGCTTTAGGGGACATGGAAGGGCAGCCCTGGGATAT TTTTAATGAGAACAGTGACTTCCTGTTTAATTCCCGAGGCTTGTCTCTTT TGCCTACCATACCCACACTGGTATCACAAGATACCGCCCATGATTGGGGA GGGGGTTCACCAGACTGGCCTAGGGAGTCCCCTGCAGGAAGCTGCCACAT GGAGAGGCTACAGCCAGCCTCACTCCCAACCCTAAGCTATTGCCCACCTT TTGCAACTCCTGAAGATTACAGCTTTCTGATCCCTTCCCCCCCCCTACAC CAGAAGGGTCCTCTGTTGTGGTCATTCAATAAATGATATTTCTTAATTAG GAATCTAGCTCTTTCTTATTCAGCTGGACTAATAAGCACCCTATGCCCTG CTTGGGTGTGATAATTTTGAGTTGGAGACAAGGAAAAAGGAGTGAATGAA AGGGAGTAAAAGTCTTCACCCACAGCACTAGATTTCAGCTATGCCCAACG TGAAAATGGAAAGGGAAAATGGAAAAAAAAAAAATTGGCCAAACACGCTT TAGGTTTGTTTTTCCCTCCTTTTGGGAGCTTTTTGCATTTTCCTCCCCAA TTTGGAAAAAAAATGAAAGAAAACAAATTTCTCTATCATTTAAATAAAAC AGACCTTTATGTCTCTAAATATAATACATCAAACAATGTTAGGAGTAACT AAATTATACATAAAGATACTTGTTTGTTAGATTGTTAAAGGCTGTTTGAA AAATAGAATTTCGCTGGTGAGGTGCCTCACACCTGTAACTCCAGCACTTT GGGAGGCTGAGGCAGGTGGGTCACCTGAGGTCAGGAGTTTGAGACCAGCC TGACCAATATGGTGAAACCCCATCTTTACTAAAAATACAAACATTAGGTG GGCTGTGATGGCACATGCCTATAATCTCAGCTACTCAGGAGGCTGAGACA GGAGAATTGCTTGAACCGGGGATGTGGAGGTTGCAGTGAGCTGAGATTGA GCCACTGCACTGCAGCCTGGGCGACAGAGGGAGACCCTGTCTCAAAAACA ACAACAAAAAATAAGAATATAATTTCACTTTTTGTCAGCCTCACATCCTC CATGGTTTTGTGTGTTTATTTTTCCAGATATTTTATACCTCCAGTTATGA CTCTGTAGAAAGATACCATCTGGGGGCCAGGCATGATGGCTCACCCCTGT AGTCTCAGCACTTTGGGAGGCTGAGGCAGGTGGATTGCCTGAGGTCAGCA GTTCAAAACCAGCCTGGCCAACATGGCGAAACCCTGTGTCTACTAAAAAT ACAAAAATTAACTGGGCATGGTGGCAGGCGCCTGTAATCTCAGTCACTCG AGAGGCTGAGGCAGGAAAATTGCTTGAACCCAGGAGGCAGAGGTTACAGT GAGCCGAGATCGCGCCATTGCACTCCAGCTGGGAGACAGAGTGAGACTCT GTCTCAAAAAAATAAATAAAGATACCATCTTGGCTTTCCCATATTATACA GATCCAGAAGAAAGACCAACTTAGGATCTCTATGCACATGATTATTTCAT ATTTTTTGGAAGAAAATAAACTAGTGTTGAATTTAAGAACATGCTCAGAA GTCATGATTTTTGAGGAAGGAGGCTATTTATTTAAATCGATATAAAGGAC CATTAGTTTTAGACCTGTAATTTATACTTGGTGAAATTTTCATGGAAAAA AAACAACAACAAAAAAACTCATTTCCCTAAATATTTTCTAGTAAAAACAT GGCTTGCTTTTTTGGTGCAAAGTCTGCCACGCTGTTTTTAAAAGCGAGGC TTACGAGACCGTGGGAGAGAGATAAGTGAACAGCCTCTTTAATAAGAGAG GCGTCCAGCGTGGCGGCGGAATGCAATACCAAAAAGTAAACAAAGAGCAT CGTGTGAAAAAGAGCAAGTTGAAATGAATCTTGCTTTTCCTATTTGAAAA ACACGCTCATGGTTGTTAGCAACTGAGTCAAGACATTTAAATCATATATA TACTTTTAGATCTTGACAGTGACCTTTTATAAGTGTACAGTGGGGATAAG AAGATGAGCAAAGCCTTGCTGCAGAAAAAGCATTTCAGTTAATTGAACAT GAAATGTGTTACCATCTGATAACATTAATAATATGTGATCGCTACTTTGT ATCTAATATGCAGTTCATTTGGTTGGAATCTAAAGCATTCTATAAATGTT AGAGTATGAATCCTGTTGCAAACCTATAAACTAAGCAGCTCTATTTTGGT GCATTTTGAAGTATCTCTGTGTTAGTTATCTATGCTGTGTAACAAATTAT CCCAAAACTTAGCAGCTTCGAACAACAAATATTTATTATCTCAGCATCAA TCAGGAATGGCTAAGCTGGGAGGTTCTAGTTCAAAGGCTCTCATTAAGTT GTAGTCAAGGCATTGGCCAGATTAACAATCATTTGAAGACCTGATGAGGG CCGGCAGATCCACTCATAAGGTGTCTAACTCACAATCCCAGCAAGTTAGT TTGAGATGTTGACAGTAAACCTCAATTCTTTTCTACACTGGTCTCTCCGT AGGGCATGGAGAGATGCCTGAGCATCCTCATGACATGGCAGCTGGCTTCC CCCAGAGCCAATGATCCATGAGACAAAGCAAAACGGAAGCTACAATATCT TTGGATGATCTAGCCTTAGAATTCACCCATCATCACTTCCTTCAGGTCCT GCTCCTTAGAAGCCGGTTACTAAGCACAGTCCACACTCGAGGAGAGGGCA ATTCGACTCCATCTTTTGAAGGAGTCTTAAAGAATTTGTGAGCATATTTT AAAGGCACCGCAATCCCCTATTTACATAAGGACAGTTGAAAATGATGGTG GCTTACCTGCTCAAGGTCAACGAACTACTGGTAAGACCCCACCTGGAAGG CGGCAGGCTTTTTTATTTATTGTAAAGCAAAACAGAAAACCCACATTCTT GAAATAACTGCACATGAATCCCAAATCTGTCTCTTTCAAATGTCCAAGAC CTTCTAAAAGTGGCAGGATGCTTTCTGTTTAGAAATGGATGAGATGGACA CTAGACTGGAAGGGTCAGCCTTTGATTAAGAGTCAGCTTTCCTCTTAATC AGCTCTGGGACCATGAGAACAAAAACACTTTTCTAAGGGATGTTTTCCTC CTTTGCAAAATATGATGGGCTAGCCGAATGGTTTCCAAAGTTGGTGGCCT TTAAGTCCTCTGGGGACTTAAAAACTCACTGATCTTGTGTTAAATCCACA ATGTCCAGGAATCTGTACCACTTAAAAGCACTTGGGGACTCTGGCGGCCT GTTTTGCAGACAGTAGGAACTGCTCGGCTACATGATTTCTCACTCTTCCA CTTTTAACATTATTTTATTTATTTTTGAGACAGAGTCTCTCTTTGTCACC CAGGCTGGAGTGCAGTGGCATGATTTTGGCAACCTCCGCCTCCTGAGTTC AAGCGATTCTCCTGCCTCAGCCTCCAAAGTAGCTGGGATTACAGGCTCCC GCCACCATGACTGGCTTTTTTTTTTGTATTTTAGTGGAGATGGGGGTTTT ACCATGTTGGCCAGGTTGGTCTGGAACTCCTGACCTCATGTGATCTGCTC ACCTCTGCCTCCCAAAGTGCTGGGATTACAGGCGTGAACCACCATGCCTG GCCAACATTATTTTAACTCTCCCCATCAGACTGGGTATGCCCATGTAAAT TGTTGGTTCTCCATCTTCACTACAATCATGAGCAGAATTTTTAAAAAATA TGATACCTAGGGCCCTCCCTAGGCAAAATATAAGTCATTCTGGGGTGGAA CTCTGGTACCATCACGGGTTGTTGGCTTGTTTTCATCAGTACATTTAAAA CTAATCATGTTTAGCCTTGTTGGCACATTAGAATCACTTGGGGAGCTTTA AAAAAGCCCACTGCCCAGCCTGTCCCCCAGGCCAATTAAATCACAATCTC GTGGGAAAACCAAGAATCAGCATTTTTTAAAGTTCCCCAAGTGATTACAA CATACAGCCAAACTGACCTATGTTTGCCACATTTGAGATAATTCTAATGC TAATTCACCTATAAGGGATTATTCAGAAAAAAATCCCAACATTTAGATGC CACAGTACTCTAAGAAAAAAAATGCATTTAAAGTGGAAGATATTACAATT TTGAAATGAAAGATATTAAAAATTAAATGGAACTAAGTTCCATTTCTGGC AATATGGTAGACTAAGTAACTTGAAAATCCTCCCATCATAAACCACCTAT AAATACTGGTCAGAATGTAATAAACACCCATTTAAATGAGCTCTCAGGAC AGTAAGCAAAGGCTCTCAGAGTCAGGAAGAAGAGGGAGATTCTAGCATGG TATGCAAGTAAGCTGAGGTTGAGCTGGTCTTAGGCAGGTTTGCTGGTGTT GGGAACCTGAGGTTTGAGCATCAAAATAGGAAGGAGACTATGCTTAAGGT CCATTAAAAGTGGGAAAATGGAATTCAGAATTCCCATAAAGCTGGAATCC CATCAAGCTAGAACCTCCTGAATCACTAGAGAAATAATCACTGGAAAAAT AATCTCCCCAATGTCACAAGGAAACAAGAAAATGTGCCTGTCTTTGCAGG GGTTGAGGGTGGGGAATAAAGGGCTTTACTGAGAATTTGAGATTATAATG TGGTATGGTCCAGGAACCCCAAAGCTGAGAATGAATACAGAAATACAGAC CCAATGCCAAACTATACAATGTATGTGGATATAATCCTCCACAAGCAAGA TGTAGCAGACACAAAGGTCCCAAGAACCTCAGGTAACAGAACTATCAGGC AGACTATAAAATAAGCAAATTGAAAATTATTAAAGACACAAAGAGGCCGG GCGCGGTGGCTCACGCTTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGG CGGATCACGAGGTCAGGAGATCGAGACCATCCTGGCTAACACGGTGAAAC CCCATCTCTACTAAAAATACAAAAAAATTAGCCGGGCGTGATGGCGGGCG CCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACC CGGGAGGCGGAGCTTGCAGTGAGCCGAGATTGCGCCGCTGCACTCCCGCC TGGGCCACAGAGCGAGACTCCGTCTCAATTAAAAAAAAAAAAAAAAGACA CAAAGGAACCTTTGACACCATGAGAAAATAACATAACACTCTAAAAAAGG TAGATTTTAAATAGAACTAAATAGAATTTCTAGAAATGACAAATATAGTC ACCAAAATGAAAAGCTCAGTGATGAGTTAAACAGCAGATTAGACAGAGTC GAAGAAAGAACCACAGATGGATCTGAGAAAATTGCCCAGGAAGCAGCAAA GACAAAGTGAAGGAAAGTCTGACAGATTCAGAGTGTGCTGGATAGAAGGA GAAGATGCATTATACATTTCATATAAGTACCAAAAGACAATGAGAGGGAT ACTTCATTTAGAGAATCCCAAGATTATGCATATACAATGAGTATTGAATC AGATAAAGAAGAAGAAATTCATACTTGAATATAGCAGAGTAAAAATGTAG GAAGCCAAAGACAAGGAAAAAGTCTTAAAACCAGAGAGAAAAGACAGATT ACCTACAAAGGAATGACAATTAGACTCATAGCAAATGTTTCAGAAATAAA GAATAGGAAGACATGGTATATTCAAAAAAGTGCTGGGGTAAAATAACTGC CAATCTTGAGTATTACACCCAGAGAAATCATCATTCAGGAATGAGAGTGA AATATGACATGTTTGTCTTAGCGGAGAGAGCGTACCACTCAACAATCCCC TGAAAAAAACTAAAGGTATGTTTCAGGGGAAAGGGTCTATATCTAGAAGG AAATTGGTAAATAAGGGCAAATCTAAACGATGAATTGACTGTATATAAAA TTACAATAGAGATTAAAATTAGGGGTATAAAAAGTAGGTGGATCTAAAAA TAAGCAACAGTAAAACATAATGAGAGGATGTAACTGAAGTTGAATCATTC TTAGCTTATTGGATAGTTCTAGGGCATTTGATTTACTTTAGATCACATGT ACAGGTTAAAATTGTAATCACCGAAAGAGTAGAAATAGAATTTACAACTT CCGGCCAGACACAGTGGCTCACGCCTGTAATCCTAGCACTTTGGGAGGCC AAGGCAGGCAGATCAATTGAGGTCAGGAGTTCAAGACCAGCTGGCCAACA TGGTGAAACCCCGTCTCTACTAAAAATACAAAAATTAGCTGGGTGTGGTG GTGGGTGCCTGTAATACCAGCTACTCGGAGGCCGAGTCAGGAGAATCGCT TGAACCCAAGAGGCAGAGGTTGTAGTGAGCTGATTGTACCACTGCACTCC AGCATGGCTGACAGAGTGAGACTCTGTCTCAAAAAAAAAAAAAAGGCCTC GGCCTCCCAAAGTGCTGGGATTACAGGTGTGAGCCACCATGCCCGGCCAA TAGCATCTCTTATACATTGCTAGTAGGAGTACAAATTGGAACACCACTTT GGAAAACAGCTTAGTATTACCTTGTAAAATTTTACATTCACGTATGTTAC GACCCAGCAATTACTCCAAAGAGAAATTCTGATCTATGTGCATCAGAAGG TAAAAATGTTCATAATAACACTGTTTATAATAGCCAAAAAAAAAAAAAAA TTCCTGAAAGCAACCCAAAGGCTTGTTTGTGAGAATAGAGAAACTAAACT GTGGCACAGTCACATAATGGAATATTATACAACTGGGAGAAAGAATGAAC TACAACCTGATACAAAAATCTAATTTTGTTCCCCCCACCCCCCCAGGGCC CTGGCTAGAGGATCTAATTGATTCTTAATAATTTCATATTGAGTAAATTA TCAAGCCTCAGAAATTTTTCATAAAGTTTCAAAACAAAAAGTAAAACAAA ACAAATATTAGTCATTGATATGCATGATAAAACTTTTTAAAAAGCGAAAA ATCATAACAAATACAAGATTCAGACTGGTGGTTACTTTAGAGGAACAAAA TAGGGAGGAACACATAAGCAGATGTTACATAAGTCAAGTCATTGTTTCTG TTTTAGTTCTCTGGTTGAATAGCAGGTTTACAGGTATTCATGATATCAGC AAATAAAATAAAATAGGGCCATGAGTATACACAATGATGATAGTGTGTTA TACTAGAGATTGTGACTAATATATTTTTGTGCCCCTAAATTGTGATATTT TGTGATCTTTAAAAATATCAGCAATCACAATAAATGTAAATGGACTAAAC TTACTAGTTAGCACAATAATGAACCAGAGCTAACTATATGCTATTTACAG GAGACTGAACAAAAAATTTGGGACATGGAAAGGTTCAAAGTAAAAGGAAA GAGGGAAAGAGAAAGATCTATAAGTGAAGGGCTAATTAAAAGAGTAGTTA CATTTATTACAGACAAAGTAGACTTTAAAGCAAAAGGCATTAGGGATAAG TATCTTCTGTGCCAATATTAAAGGAATAATTCCTAGGAAAATAGCAAATC TAAGCTCGTATGCACTGAATAACAAGGCCTTAAAATACACAAAGCAAAAA GTATGAGAAGCAGAAAATGTATAGTTACAGTGGGAAATTTTAACAGTTCT TTTTGATGCACCTGACAACATACTTTTAAAATACAGCAAAATTTGAGAGA ATTACAGGAGAAACTGGCAAATCTAGTCATATAGAAATTTTAATGTACTT CTATAAAAACCAACAGGTCAAGCTGACAAAATATTAGTAAGGCTACAGGA AATCTAGACAATATAATTAATAAGTCTGATCTAATAGACCCATATAAAAT GCTGTACTTAAAAATTAAGGCATATACATTCTTTTCAAATCTACATGAAA TATTTATAAAAATTGACTGCACCAAAGCAAGTTTCAATAAATAACAAAGA ATCTGCTTCATATAGATCACATTCTTTGACCTTAATTCAGTTAAGTTAGA CGTTAGTAACTAAAGAATAACCTCAAATACCCCTCAAATACACCTGTATT AATTCACTAGTCAAACAAGAAATCGTATCTGAAATCTAATTTTTTTTTTT TTTTAAACAGGATCTCGCTCTGTCACCCAGGCTGGAGTAGTGACATGATC ATGGCTCGCTACAGTCTCAACTTCCCAGGCTCAAGTGATCCTCCCACCTC AGTCTCCTGAGTAGCTGGGCCTATAGGTGCATGTTAGCACACCTGGCTAG TTTCTAGAAAAGTTTTCTTTTGCAGAGATGGGGCCTCACTATGTTGCCCA GGCTGGTCTCAAACTCCTGGGCTCAAGTGATCCTCCCACCTTGGCCTCCC AAAGTGCTGGGATTACAGGCATAAGCCATTGTGTCCAGCCTAAAATCTAA ACTTTTTTTACCTCATTGGTGATAAAAAGTCTATATATCTTGTGGAATAT AGCTTGAGTGGTACTTAAAGTGAAATCTGTAGTCTTAAATGTTAAAATCA GAAAACAGATGGCTAAAAATTAATTACCTAAGGCAACAATTCAAAAAAAA AAAAAAAGAGCAACAGAATAAATCCAAAAAGAGCAGAAGCAGAAGGGAAT ACTATATGATATATATTAATATTACATTACATATTATATTCTCACAAACT ATATAATAGTATATATTTTCTATTGTGCCATATATAACAATATAATATAT AATAATATATAAGAACAAAACTTACTGGAACAGAAAAAGAAACAATATTG AAACTAAAGTCTGGTTCTTTGGAAACACTAATAGAAAGTAGAAAGAAAAC ATTAGTAAAAAAAAAAGATCTCTGGCACAGCTGATCCAGATAAGAAAAAA TACATATAAAATGATATTTAGAATGAAAAAAAAACATTATGGATAGATAG AGCAGAGATTAAAATACATTAAGAGATAACTATGAACAGCATTTTGTCAA CACATTTGAAAACCTAGATGGTATGGATAATTTCCTTGAAAAACATAGTG TATAAAAATAGATTCAAGAAGAAAACAGAAAACCTGAAGAGATCTACGAG CATTAAAGAAATGGAATCAGTAGTTAAAAATCAACCCACGAAATGTCTAT CCCAGACCCAGACAATTTACCTGCAAAACTACCAAACATTCAAGGAACAC ATAATTCCAATCTTACACACACTGTTCCAGAAAATGAAAAAAGAAGGAAC ATTTACCGGTTCATTTTATGAGACCAGTATAACCAGACAAAGGCAGTAAG AAAAGGAAAAACTGCAGCCAATTTGACTTATGAACATGAATACAAAAAGA ATCCTAGAATGAAATTAAATGTTGGCCATCATTCATTCATTCATCCACTC ACTCATTCAGTCAATCATTATTTATTGAGCGTCAACCGTGCGCCAGCAGG CACTGTGCTAGTACATGGAGAGCAGAAAGGCACGGGAGCTTCTGGCTTAG AGGAGATGGGCAATAAAGCAAATGATCATACAGGGTAAGGTACACAGAGG ACGTTCTGGTAAGGTAACTGCATATCAAAGGGCATTCGACCCTGTCAGAG AGGTCTGGGAAAGATTTCCAGGCATGTAAGTGGAGTAAGGGTGTATGTGG GAAGACTGTTTTGTAAGCTGTTGCAGGGCCTCAGGTGGGAGATCTGGGAT GCAGCAGCAAGAAAGATGGATTTGAACTTGGGCTTCCTTTAGAAAGGCTA AGTGGAGATGTTGAATAGGAAATTGACCAGAGCCTGGAGCTCTTCAGGAA GGGTGGGGCTGGAGATTTCAATTTGAGTGGCATCACCATGTGTTTAAACC CATCCTGGAAGATTGAGTTTGAAGAAGGAAGTGTCCAACATTGTCTTGGG CTGTTGAGACTTTCAGAGGGTTGAGGACTGATATTGTGCTGCTTGAATTC TCCTGATGCAGGGGCTACATTGAGTGAGCTGGAGAAAAAAAATGCATAAA ATAATAATAATAATAATAATAATAATAATAATAAGCTATTACAAATAATG TAGAGCAAAGGGGCAGCAAGAGGGAATTTTTTTGGACAATGAAACTGTTC TGCATCTTGATTATGGTGGTGGTTACATGACTCTATACATATGTCAAAAC TCATAGGATTACAGACCAAAAAGAGTAAATATTACCGTATATAAATTAAA AATAAATGAGTAAAAACAATGTAGTAATGGAGACTTAAAATCCAGTTCTT TCTAAGCCCTGACTTTGTAACCGCAGCTCTAGCCCCTCTCTGGATTTTAA ATCAGTTCTATAAGTGTCAGCTTGTGGAGGTCTATACCAGACAGGAAGGG CCCCCAACTCTCGCCTTGTGAGGGACAGAATAAACACGCAGGCAGCAGAG GCCACACGGCATTGGACTGATGGTCAGAGGGTGGGGGTGGGGTGTAGCCT GGTGAGTTTGGCACCTCTGAGACGCTGATGTATAATGAGGGGATTAGATT AGGAAAGGCCTTTCTACCTAGGATGGCCTGTGGTTCTACTGTAAAAATCC CAAACACAATACAATTAGCTCTGTTGTCTGCATTTTGTTTAGAATAATCA ATCATAATAAACAATCATTGTAACAACTGGCTGTTCAACACATGAGACCC CAGATGATTTGGGAAGGAGCTTGGAGTGACAGGAAATGTTTGGGTTTGTG GTTTAAAGCCTTAGAGCACCTTCTCAATATGATTATATTGAGTAGTGATT GATAATAAACACGACTCAGGTTTACAGTGAAAAAGGAACTTTTACAACAT TGGTTCACTTCAGCCTCTCACCTTCACCACATCAATCCTGTCAAGGAGGA ATTACTGCAATTTAGGGAACAGGGAGACTGAGGGTCTGGTCACTCAAGGC TATGGCTGGTGTTGAGATTTTCCCAATATTCCATTTTTCCAAAGCCCACA GTGGATTTGGTTCAGTTTTGGTGTTGAGTGTATTCCTTTGTCTCCTAATC CTATGAAAATTAATGGAAAAGTGTTAATTGGGCATCAATTCATGCTTAAC ATTAATCTCAGTATTTGATGAACCACAACTTTATGTTGCCCCTCATGCCA TATTAACTCAGTTTATTGCAACAATTTAAAACGATACAGATTTAAAACAA TATGGGTAATTGTATCCGTATTGTTTCAAATGCCCCATAAATTGAAACCA GCCCGAATTTGGGCAGTCTGGAATCTGCCGGAGAAACTTTCATGCGATGC CTTTGGAAGGCTACAGACATTGTCTTTTTGGAGTTTTCAGTGCATGAAGG TATGAAACCGCATTTATTAAGCACCTACTGTATGCCAGAACCCGTGCTGC ACAATACTACTGCTGCTAAGGTGGGAGTGATTCTGAAGCCTTCTGCCACC CTAGCTACCTCTGCAGGTCGTGAGGGGTCTTGGGCTATTTCAGTATCATG CACTTTACTATCCTGGCATACAAAGGCTGGGTGAGAAATAAAATATATAA CGAACGGATTACACAGGGGTTTCCTGAAATAACCACCCTTCCCATCCATC CCAGAGACACCCCAAAAGTACTTCTCGTTATATACAAACATTTGCTTTGA ACCTCAATCATGTGACCTTGACTCCTATAACCTATCTTATTACATTTTTA AAACACTGTATGATTAACGCGGAAACCCTTTCTTCGGCACTTTCTCGCCA CTGGAATCGCGTCAGTTTCTCAAAGTTCCAAAATAACCTTTCCCGGGCAC GGATTGGTACCTCTACTGGGGAAGGGCGGGGAACCGCGCAAGACGTGCCG GTGTGGAGCCAGAGCCAGAGAGAACTTCCAGCGCAAAAGGAAAATAAAAC TTGTGGCTGGTGTTTGTGCAGGAGGGTCTCCGCCATCCTGAAGCCCCCCG ATCCTGGGGCGTCTCGGGGGCCGCCAAAGGAGCGCCAGGGTGTGGGTTTG CTCCCGACGTCCTTGACCTAAATTTCTGAGCGGTGGCTGGAAACAGGGCA CAGCGGAGGGCGGGCGGCTGGTGCCATTCCCGGATCTCGGCGGCAGGGGC CGGCAAACTTGAATGGAGAGGGCGAACTAGAGAGGGTGGGGGGCGTCTTC TCCCAGGTCCGGGTGAGGAGCCGCAGCAAGCTCCCCGCGCCTCCCCTCCC CCGATCCACCCGCCCCCCGCAGCCCATGTGATCCAGGGAAGTCGGGGTGC GCTCCCCCTCGCCCTGCGCCCTGCCGGCCCGGAGGCGGGGTCCCCTCCGC CCGCGGGGTTCGCGCGCCACCCTTGTGGGTCCGGCCGTGGGGGGCCGAGT GTGCGCGCGCGGGCAGGCGGGGGCCGCACGGGGGTGCGTGACGTCACCGG CATTGGTTACACGACGTTCTAGAACTCCGCCCCACGTGCGCCGGGGAGGA GGGGGAGGAGGAGGAGGAGATGGGGGTGGGGAGGAGGAGGGGGAGAGGTG GGGATGGGCCGGGGGGGCGGGGACGGGGGGGTGTGCGAGGCAGCGGGGCT GAGCTAAGCCGAGCCCACGTGTGACGGCTCTCGCCGCTGCCCCGGCTCCG CCGCTCGCAGAGAGATTCGGAGGAGCCCGGGCGGGGGGGAGGAGGAGGGG GAGGAGGGAGCGGAGATCTCGGGGCTCGGAGCCGGCCGCCGCTCCGCTCC GATCGCTGTGGGGCTTGGTTTTTTGGGGGTGGGGGGGCGGGGGGGCTCAG AT ATG

JAK2

The JAK2 gene is located on Chromosome 9. JAK2 protein promotes the growth and division (proliferation) of cells and is part of the JAK/STAT signaling pathway important in transmitting signals from the cell surface to the nuclei. JAK2 is especially important for controlling the production of blood cells from hematopoietic stem cells. These stem cells are located within the bone marrow and have the potential to develop into red blood cells, white blood cells, and platelets. Essential thrombocythemia is characterized by an increased number of platelets, with the most common mutation being V617F found in approximately half of the affected people, with a small proportion having a mutation in exon 12. The V617F JAK2 gene mutation results constitutively activated JAK2 leading to the overproduction of megakaryocytes, and hence excess platelets. As a result, there is increased risk of blood clots and decreased availability of oxygen. Overproduction is also associated with primary myelofibrosis, as megakaryocytes stimulate other cells to secrete collagen thereby replacing bone marrow by scar tissue. The V617F mutation is found in approximately half of individuals with primary myelofibrosis. A small number of people with this condition have mutations in the exon 12 region of the gene. These JAK2 gene mutations result in a constitutively active JAK2 protein, which leads to the overproduction of abnormal megakaryocytes. These megakaryocytes stimulate other cells to release collagen, a protein that normally provides structural support for the cells in the bone marrow but causes scar tissue formation in primary myelofibrosis. The V617F mutation is occasionally associated with leukemia, other bone marrow disorders and Budd-Chiari syndrome.

Protein: JAK2 Gene: JAK2 (Homo sapiens, chromosome 9, 4985245-5129948 [NCBI Reference Sequence: NC_(—)000009.12]; start site location: 57256743; strand: positive)

Gene Identification GeneID 3717 HGNC 6192 HPRD 00993 MIM 147796

Targeted Sequences Relative upstream location to Sequence Design gene start ID ID Sequence (5′-3′) site 12063 CGCACCAGTTTGTCCACGTCCAG 1663 TG 12098 GCCGTCACTGCCGACATAAGCACA 1811 GAC

Target Shift Sequences Relative upstream location to gene Sequence ID Sequence (5′-3′) start site 12063 CGCACCAGTTTGTCCACGTCCAGTG 1663 12064 GCACCAGTTTGTCCACGTCC 1664 12065 CACCAGTTTGTCCACGTCCA 1665 12066 ACCAGTTTGTCCACGTCCAG 1666 12067 CCAGTTTGTCCACGTCCAGT 1667 12068 CAGTTTGTCCACGTCCAGTG 1668 12069 AGTTTGTCCACGTCCAGTGT 1669 12070 GTTTGTCCACGTCCAGTGTC 1670 12071 TTTGTCCACGTCCAGTGTCA 1671 12072 TTGTCCACGTCCAGTGTCAA 1672 12073 TGTCCACGTCCAGTGTCAAC 1673 12074 GTCCACGTCCAGTGTCAACT 1674 12075 TCCACGTCCAGTGTCAACTG 1675 12076 CCACGTCCAGTGTCAACTGA 1676 12077 CACGTCCAGTGTCAACTGAG 1677 12078 ACGTCCAGTGTCAACTGAGC 1678 12079 CGTCCAGTGTCAACTGAGCA 1679 12080 TCGCACCAGTTTGTCCACGT 1662 12081 ATCGCACCAGTTTGTCCACG 1661 12082 GATCGCACCAGTTTGTCCAC 1660 12083 GGATCGCACCAGTTTGTCCA 1659 12084 GGGATCGCACCAGTTTGTCC 1658 12085 TGGGATCGCACCAGTTTGTC 1657 12086 TTGGGATCGCACCAGTTTGT 1656 12087 CTTGGGATCGCACCAGTTTG 1655 12088 CCTTGGGATCGCACCAGTTT 1654 12089 GCCTTGGGATCGCACCAGTT 1653 12090 GGCCTTGGGATCGCACCAGT 1652 12091 GGGCCTTGGGATCGCACCAG 1651 12092 GGGGCCTTGGGATCGCACCA 1650 12093 GGGGGCCTTGGGATCGCACC 1649 12094 TGGGGGCCTTGGGATCGCAC 1648 12095 CTGGGGGCCTTGGGATCGCA 1647 12096 TCTGGGGGCCTTGGGATCGC 1646 12097 ATCTGGGGGCCTTGGGATCG 1645 12098 GCCGTCACTGCCGACATAAGCACAGAC 1811 12099 CCGTCACTGCCGACATAAGC 1812 12100 CGTCACTGCCGACATAAGCA 1813 12101 GTCACTGCCGACATAAGCAC 1814 12102 TCACTGCCGACATAAGCACA 1815 12103 CACTGCCGACATAAGCACAG 1816 12104 ACTGCCGACATAAGCACAGA 1817 12105 CTGCCGACATAAGCACAGAC 1818 12106 TGCCGACATAAGCACAGACA 1819 12107 GCCGACATAAGCACAGACAA 1820 12108 CCGACATAAGCACAGACAAC 1821 12109 CGACATAAGCACAGACAACT 1822 12110 CGCCGTCACTGCCGACATAA 1810 12111 TCGCCGTCACTGCCGACATA 1809 12112 ATCGCCGTCACTGCCGACAT 1808 12113 AATCGCCGTCACTGCCGACA 1807 12114 CAATCGCCGTCACTGCCGAC 1806 12115 CCAATCGCCGTCACTGCCGA 1805 12116 GCCAATCGCCGTCACTGCCG 1804 12117 AGCCAATCGCCGTCACTGCC 1803 12118 CAGCCAATCGCCGTCACTGC 1802 12119 CCAGCCAATCGCCGTCACTG 1801 12120 CCCAGCCAATCGCCGTCACT 1800 12121 ACCCAGCCAATCGCCGTCAC 1799 12122 TACCCAGCCAATCGCCGTCA 1798 12123 CTACCCAGCCAATCGCCGTC 1797 12124 CCTACCCAGCCAATCGCCGT 1796 12125 GCCTACCCAGCCAATCGCCG 1795 12126 TGCCTACCCAGCCAATCGCC 1794 12127 TTGCCTACCCAGCCAATCGC 1793 12128 CTTGCCTACCCAGCCAATCG 1792

Hot Zones (Relative upstream location to gene start site) 1550-1900

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13675) GTCATTTATTTCTGCTGTGAACTTCATTTTTTCCTTCCTTCTGTTAGCTT TGGGCTTTGTTCTTCTTTTTCTAGTTCCTTGAGGTGTAATGTAATGTTGT TTGACATCTTTCTTCCTTTTTGATGTAGGTATTTATTGCTATAAACTTCC CTCTTATAACTGCTTTTGCTGCATTTAATACTGACTATAATAAGATACGA TGTAATAGATTTCAAGGAATTATGTATTTTTGAATAAATTAATTCTTTAA AGTTGCATATCCAGTTGCAGATGAACTTCAAAAATCTTGCAGTTTTATAT CTGTTACAGTAATTGCCAGGTTTTGTTGTTGTTGTTTTGATACATTAGAA GTTCTAGAATTGTTATATCCTCTTGATGAATTAATCCCTTTATCATTCTA GAATTACCTTGTCTCTTTACTGTTTGTGACTTAAAGTCTGTTGTATCTGA TATACCTTTGCATGGAATATCTTTTTCTATCCCTTTACTTTCAGTCTATG TGTATCTTTAAAGGTGAGATGAGGTTTTGTAAGTGGCATGTAGTTGGGTC ATGTTTTTTAGTCCATTTAGCCATTCTCTATCTTTTAAGTGGAAAGTTTA ATCTATTTACATTCAAGTTTATTCTTGATATGTGAAGGCTTATTCCTGTC ATTTTATTAATTGATTTCTGGTTGTTCTGTAGGTCCTTTGTTCTTTTCTT TCTCTCATATTGTTTAGCATTGTGGTTTGTTGGTTTTCTATAGTGATAAC ATTTGAATCCTTTCTTGTCTGTGTGTGTTTGCTTTACCAGTGGGTTTGAT ACTTTCGTCATCTGTTTTTCATAATGGTAGTAATTGTCCTTTTTGTTTGT TTGTTTGTTTCTTTTTTGAGACAGGGTTTTGCTCTTGTTCTGTCCTCCAG GCTGGAGTGCAGTGGTGTGATCATGGCTCACTGCAGCCTCGACCTCCATG GTCTCAGGTGATCCTTCTGCCTCAGCCTCTCAGGTAGCTGGGACTACAGA AACCTGCCACCATGCCTGGCTAATTCTTTTGTATTTTTCGTAGACATGGG GTTTTGCCATGTTGTCCAGGCTGCTCTTGAACTCCTGGGCTCAAGCAGTC TGCCTGCCTCAGCCACCCAAAGTGCTAGGATTACAGGCTTGAGCCACTGT GCCTGGCCTGACATTGTTCTTTGACTTCCATATGTAGAACTCCCTCAAGC ATTTCTTGTAGGTCTGGTCTAGTAGTGTTGAATTCCTCAGCTTTTGCTTG CCTCAGAAAAACTATTTTTCCTTTGCTTAATGAAGGATAATTTTGCTGGG TATAGTATCCTTGACTTGCAGGTTTTTTTCTTTCAGCACTTTTCATATAT CGTTCCATTCTCTTCCTGGCCTGTAATGATTCTGCTGAGAAATCTGCTGT TAGTCTGATGGAGCTTCCCTTAGAAGTGACTAGACTCTTTTTTCTTGCTG TTTTTAGAATTCTCTCTTTGTCTTTGACAAGCTGTTGTCTCTGACAACAG TTCTCTCTTTGTCTTTGACAAACTGTTGACAGTTTGACTCTAATGTGTTG TGGAGAACCTGTTGGAATTTTGTCTTTTTGGGGATCTCTGAGCTTCTGTA TCTGAATGTCTAAATCTCTTGATATACTTGGGTAGTTTTCAGCTATTATT TCATTAACCAGGTTTTCTATTCCTTTTGTATTTTCATTGTCTTCTAGAAT ACTGAAAATTCTAATATTAGTTTGCTTTATGGTATCCCATATGTCATGCA GGCTTTGTTCATTCTTTTTTCTTTATTTTTGTCTAATGGGGTTATTTCAG AAGACCTGTCTTCAAGTTCAGAAATTCTTTCTTCGTAGATGCTCTAGAAT GTATTTTTTATTTCATTAAATGAATTCTTCAGTTTCAGGGTTTCTTGTTT TCTTTTTAAATGATATCTCTCTCTTTGGTAAATTTCTCATTGATATCCTG AGTTGTTTTTCTGGTTTCTTTGTATTGTTTATCTGTATGCGTTTGTATCT CCCTGAGCTTCTTTAATATCATTATTTTTAATTCTTTTTCTGGCATTTCA TGAATTTCTTTTGCATTGGAATCTTTTGGTAGAAAATTATTTTGATCCTT TGGAGATGTCATATTTCCCTATGTTCCCATGTTTCTTGTGACCTTACTTC TTTGATATCCACACATCTGGTGTAATCATCACTTCCATTTTTTTGAATTT GCTTTCATAGGGTAGGACTTTTTCCTGAAGATTTGACTGGGGTGTTTGTT GGCCAGGGCACTTTGGGTTTGAATCTGGGTGCATGCAGTAGTGTAGTCTC TGTAAGATTTTTTTTCCTTTGTAAACAGCATCAGTGGTGTCTGTGATTTC CTCAGTGGCATAGTGTGTGGTTGTGGAGGCTGTGGTGAACTTTTGCTGGG GATGGTGACACCAGCTGGACTGATCCTCAGTCCTCAGTTGTGGCAGCAGT TGGACAACCATGCCTGTACATTAGCCCCAGGGTGGCTTACATTAGTAATG GTGTTAGTGGGTCCAGGCAGTCCAATTTTTGGGTCTCCAGGTGACTTGTT TGGGTACCAGGAGTGGCAGTGATGGGCTGGGCAGCTGAGTGGGTCCACAG GCCCCTGGGCAGTGAGCATGGCATGGGTTATGTCAGTAGCAGTGGTAGGA GAACCTCTGGCTGTCCAGTTGTCTGTGCTTATGTCGGCAGTGACGGCGAT TGGCTGGGTAGGCAAGTCCTAAAACCTGCAGGTGGCAAGTGTGAGTGGGA ACCAGCTGTGGTGGTAGTGGCAGGTTGGGTGGGCCACATCCTCAGACCCC CAGGTGGAATGCTCAGTTGACACTGGACGTGGACAAACTGGTGCGATCCC AAGGCCCCCAGATAACATGCTTGGATACGTGGGAGTGGGGTGCTGAGCTG GGCAGGGTGAGAGTATCCTCAGGCCCTCCAGTGGTGTTAGCAGGTGCTGT TTGTGGTGGGCAGGAGCAGGATGATTTCCAATTTCCTGGTGGAATGTTCA GGTGGGGGCAGCAGTGGCTGTGCTGTGCCCTGATGCTGGGGAGGGTGCAG TTGCTGTCAGTGGGAGCAGTTGTAGGGAGTTGGCTAAGGAGTGTGCACTG CAGCTGCAGGTGGAGGCTGTAGATGTGATGAAGCTGTACTCAGGGTGCAT GCAAATTTGCATTTTGACACCTAGCGGCAGCAGCCTGCAATGGTGGCAGC TGTAGGTGGTAGAGCTTGTCCTCAGGGCACATACCAATATATGGCAGCCC TTCTGCTGGGAGCAGTGGGGTTATTGCCAATGGCTTGTGCTTTGGTCCCA GAGGCGGCAGCCAGCAATGGAGGTGACTGTCGGTGGAGGATGTCAGTGGG GCTCTAGGGGTGTGGATATGCAGGGGCTGTTGGGCTCCAGGGTAGGAGGC ATTCTGGTGTGGGTTGGGCTTTAAAAATGGCACCGTGCTGTAGCTGCTTA GGACTCAGGGGTGTGTTGGACCAGCATAAGCTCCCTCTCTAAAGCAATGT CATTGTGCAGTCTCCAGGCAGCTCCCTATGTTACTCCCAGGGCCCATGAA AGTTGACGGGCTCTCTTGTGTCTGGGATTGCAGGAGTTTGCAGTGAAAAT GTGGGCCACTGGGAGTCTCTCACTTACTCTTTCCCCACATTGTGCAGGCT CTCTAGGCTTCTGGCTGATCCTGGCTGAGCAGGCTGCCCCACTTCCCTCT CCTTCCTTGCATTAGGTGTTTTCTATCACTTCTCTGTTGAATTTCCGTGT TCTCTCTTAGATGACCTATTCAAAGTGTGATTATCTACTCGCTATTTTGG TTCTTCTTTGTGGAGCAGGTGAGTACCAGATAACTCTAGTCAACCTTCTG GACCCCTCTTCCCCCAATTTGAGATCTCTTCTTCTGTTGTCTGTAACTGA GTTTAATGCTTGTTTGTTCATGTTAGGATTTTATATCATCGTCCTCAATT AGGTTGTTAACTGGAATTTTATAATCTTTGTCCACAGGAAGTTTAAAATG TATGATTTCTTGCATTGTGCTTTGTATGTAGTAATACACGATATTTATCC AGTTAATGGATTTGACAGCCATTGCTGTCAAGGAGCAGTCCTTCTTTGTG TATGAAGGGTGCCTTATCAATATTATTTCCATTTGTAACTTTATTTATTT ATGTATTCATTTTTGAGACAGGGTCTTGCTGTGTCACCCAGACTGGAGTG CGGTGGAGTGCGGAGGTTTGCTGCAGCCTCATCCTCCCAGGTTCAAGCAA TTCTTCCGCTCCACTCCCAGAGTAGCTAGGACTACAAGTGCGTGCTGCCA CGCCCAGCTAATTTTTTTCTTTTGTATGTTTTTGTAGAGATGAGGTTTCA CCATGTTGCTGAGGCTTGTCTCCAACTTCTGGGCTCAAGCTATCTGCCCG CCTCGGCCCCGCAAAGTGCTAGGATTACAGGTGTGAGACACTGCGCCCAG CCCATTTGTAACTTTATTGTTTTCTCTTACAGGCAAATGTTCTGAAAAAG ACTCTGC ATG

CCND1 (Cyclin D1)

Cyclin D1 belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance throughout the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin forms a complex with and functions as a regulatory subunit of CDK4 or CDK6, which are required for cell cycle G1/S transition. Regulatory component of the cyclin D1-CDK4 complex is believed to phosphorylates/interact and inhibit tumor suppressor retinoblastoma protein, RB1 to regulate cell-cycle during G1/S transition as phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complex and the subsequent transcription of E2F target genes which are responsible for the progression through the G1 phase. Further, CCND1 expression is believed to be regulated positively by Rb. Mutations, amplification and overexpression of CCND1 alters cell cycle progression and are observed frequently in a variety of tumors including mantle cell lymphoma (characterized by the t(11; 14) rearrangement) and other B-cell lymphomas.

Protein: Cyclin D1 Gene: CCND1 (Homo sapiens, chromosome 11, 69455873-69469242 [NCBI Reference Sequence: NC_(—)000009.12]; start site location: 69456082; strand: positive)

Gene Identification GeneID 595 HGNC 1582 HPRD 01346 MIM 168461

Targeted Sequences Relative upstream location to gene Sequence Design  start ID No: ID Sequence (5′-3′) site 12129 CGCTGCTACTGCGCCGACAGCCCTC 133 12242 CGGCAGAATGGGCGCATTTCCAAGA 612 12287 ACGCCACGAGGGCACCCACGGGCGGA 637 12332 CGGTGACCGCGGCCTGGGCGGATGG 2755 12388 CGGGACTCAGCGCGGCTGCGCGCCG 2907

Targeted Shift Sequences Relative Sequence upstream ID location to gene No: Sequence (5′-3′) start site 12129 CGCTGCTACTGCGCCGACAGCCCTC 133 12130 GCTGCTACTGCGCCGACAGC 134 12131 CTGCTACTGCGCCGACAGCC 135 12132 TGCTACTGCGCCGACAGCCC 136 12133 GCTACTGCGCCGACAGCCCT 137 12134 CTACTGCGCCGACAGCCCTC 138 12135 TACTGCGCCGACAGCCCTCT 139 12136 ACTGCGCCGACAGCCCTCTG 140 12137 CTGCGCCGACAGCCCTCTGG 141 12138 TGCGCCGACAGCCCTCTGGA 142 12139 GCGCCGACAGCCCTCTGGAG 143 12140 CGCCGACAGCCCTCTGGAGG 144 12141 GCCGACAGCCCTCTGGAGGC 145 12142 CCGACAGCCCTCTGGAGGCT 146 12143 CGACAGCCCTCTGGAGGCTC 147 12144 TCGCTGCTACTGCGCCGACA 132 12145 CTCGCTGCTACTGCGCCGAC 131 12146 GCTCGCTGCTACTGCGCCGA 130 12147 TGCTCGCTGCTACTGCGCCG 129 12148 CTGCTCGCTGCTACTGCGCC 128 12149 GCTGCTCGCTGCTACTGCGC 127 12150 TGCTGCTCGCTGCTACTGCG 126 12151 CTGCTGCTCGCTGCTACTGC 125 12152 TCTGCTGCTCGCTGCTACTG 124 12153 CTCTGCTGCTCGCTGCTACT 123 12154 ACTCTGCTGCTCGCTGCTAC 122 12155 GACTCTGCTGCTCGCTGCTA 121 12156 GGACTCTGCTGCTCGCTGCT 120 12157 CGGACTCTGCTGCTCGCTGC 119 12158 GCGGACTCTGCTGCTCGCTG 118 12159 TGCGGACTCTGCTGCTCGCT 117 12160 GTGCGGACTCTGCTGCTCGC 116 12161 CGTGCGGACTCTGCTGCTCG 115 12162 GCGTGCGGACTCTGCTGCTC 114 12163 AGCGTGCGGACTCTGCTGCT 113 12164 GAGCGTGCGGACTCTGCTGC 112 12165 GGAGCGTGCGGACTCTGCTG 111 12166 CGGAGCGTGCGGACTCTGCT 110 12167 CCGGAGCGTGCGGACTCTGC 109 12168 GCCGGAGCGTGCGGACTCTG 108 12169 CGCCGGAGCGTGCGGACTCT 107 12170 TCGCCGGAGCGTGCGGACTC 106 12171 CTCGCCGGAGCGTGCGGACT 105 12172 CCTCGCCGGAGCGTGCGGAC 104 12173 CCCTCGCCGGAGCGTGCGGA 103 12174 CCCCTCGCCGGAGCGTGCGG 102 12175 GCCCCTCGCCGGAGCGTGCG 101 12176 TGCCCCTCGCCGGAGCGTGC 100 12177 CTGCCCCTCGCCGGAGCGTG 99 12178 TCTGCCCCTCGCCGGAGCGT 98 12179 TTCTGCCCCTCGCCGGAGCG 97 12180 CTTCTGCCCCTCGCCGGAGC 96 12181 TCTTCTGCCCCTCGCCGGAG 95 12182 CTCTTCTGCCCCTCGCCGGA 94 12183 GCTCTTCTGCCCCTCGCCGG 93 12184 CGCTCTTCTGCCCCTCGCCG 92 12185 GCGCTCTTCTGCCCCTCGCC 91 12186 CGCGCTCTTCTGCCCCTCGC 90 12187 TCGCGCTCTTCTGCCCCTCG 89 12188 CTCGCGCTCTTCTGCCCCTC 88 12189 CCTCGCGCTCTTCTGCCCCT 87 12190 CCCTCGCGCTCTTCTGCCCC 86 12191 TCCCTCGCGCTCTTCTGCCC 85 12192 CTCCCTCGCGCTCTTCTGCC 84 12193 GCTCCCTCGCGCTCTTCTGC 83 12194 CGCTCCCTCGCGCTCTTCTG 82 12195 GCGCTCCCTCGCGCTCTTCT 81 12196 CGCGCTCCCTCGCGCTCTTC 80 12197 CCGCGCTCCCTCGCGCTCTT 79 12198 CCCGCGCTCCCTCGCGCTCT 78 12199 CCCCGCGCTCCCTCGCGCTC 77 12200 GCCCCGCGCTCCCTCGCGCT 76 12201 TGCCCCGCGCTCCCTCGCGC 75 12202 CTGCCCCGCGCTCCCTCGCG 74 12203 GCTGCCCCGCGCTCCCTCGC 73 12204 TGCTGCCCCGCGCTCCCTCG 72 12205 CTGCTGCCCCGCGCTCCCTC 71 12206 TCTGCTGCCCCGCGCTCCCT 70 12207 TTCTGCTGCCCCGCGCTCCC 69 12208 CTTCTGCTGCCCCGCGCTCC 68 12209 GCTTCTGCTGCCCCGCGCTC 67 12210 CGCTTCTGCTGCCCCGCGCT 66 12211 TCGCTTCTGCTGCCCCGCGC 65 12212 CTCGCTTCTGCTGCCCCGCG 64 12213 TCTCGCTTCTGCTGCCCCGC 63 12214 CTCTCGCTTCTGCTGCCCCG 62 12215 GCTCTCGCTTCTGCTGCCCC 61 12216 GGCTCTCGCTTCTGCTGCCC 60 12217 CGGCTCTCGCTTCTGCTGCC 59 12218 TCGGCTCTCGCTTCTGCTGC 58 12219 CTCGGCTCTCGCTTCTGCTG 57 12220 GCTCGGCTCTCGCTTCTGCT 56 12221 CGCTCGGCTCTCGCTTCTGC 55 12222 GCGCTCGGCTCTCGCTTCTG 54 12223 CGCGCTCGGCTCTCGCTTCT 53 12224 CCGCGCTCGGCTCTCGCTTC 52 12225 TCCGCGCTCGGCTCTCGCTT 51 12226 GTCCGCGCTCGGCTCTCGCT 50 12227 GGTCCGCGCTCGGCTCTCGC 49 12228 GGGTCCGCGCTCGGCTCTCG 48 12229 TGGGTCCGCGCTCGGCTCTC 47 12230 CTGGGTCCGCGCTCGGCTCT 46 12231 GCTGGGTCCGCGCTCGGCTC 45 12232 GGCTGGGTCCGCGCTCGGCT 44 12233 TGGCTGGGTCCGCGCTCGGC 43 12234 CTGGCTGGGTCCGCGCTCGG 42 12235 CCTGGCTGGGTCCGCGCTCG 41 12236 TCCTGGCTGGGTCCGCGCTC 40 12237 GTCCTGGCTGGGTCCGCGCT 39 12238 GGTCCTGGCTGGGTCCGCGC 38 12239 GGGTCCTGGCTGGGTCCGCG 37 12240 TGGGTCCTGGCTGGGTCCGC 36 12241 GTGGGTCCTGGCTGGGTCCG 35 12242 CGGCAGAATGGGCGCATTTCCAAGA 612 12243 GGCAGAATGGGCGCATTTCC 613 12244 GCAGAATGGGCGCATTTCCA 614 12245 CAGAATGGGCGCATTTCCAA 615 12246 AGAATGGGCGCATTTCCAAG 616 12247 GAATGGGCGCATTTCCAAGA 617 12248 AATGGGCGCATTTCCAAGAA 618 12249 ATGGGCGCATTTCCAAGAAC 619 12250 TGGGCGCATTTCCAAGAACG 620 12251 GGGCGCATTTCCAAGAACGC 621 12252 GGCGCATTTCCAAGAACGCC 622 12253 GCGCATTTCCAAGAACGCCA 623 12254 CGCATTTCCAAGAACGCCAC 624 12255 GCATTTCCAAGAACGCCACG 625 12256 CATTTCCAAGAACGCCACGA 626 12257 ATTTCCAAGAACGCCACGAG 627 12258 TTTCCAAGAACGCCACGAGG 628 12259 TTCCAAGAACGCCACGAGGG 629 12260 TCCAAGAACGCCACGAGGGC 630 12261 CCAAGAACGCCACGAGGGCA 631 12262 CAAGAACGCCACGAGGGCAC 632 12263 AAGAACGCCACGAGGGCACC 633 12264 AGAACGCCACGAGGGCACCC 634 12265 GAACGCCACGAGGGCACCCA 635 12266 AACGCCACGAGGGCACCCAC 636 12267 ACGCCACGAGGGCACCCACG 637 12268 CGCCACGAGGGCACCCACGG 638 12269 GCCACGAGGGCACCCACGGG 639 12270 CCACGAGGGCACCCACGGGC 640 12271 CACGAGGGCACCCACGGGCG 641 12272 ACGAGGGCACCCACGGGCGG 642 12273 CGAGGGCACCCACGGGCGGA 643 12274 GAGGGCACCCACGGGCGGAC 644 12275 AGGGCACCCACGGGCGGACA 645 12276 GGGCACCCACGGGCGGACAG 646 12277 GGCACCCACGGGCGGACAGA 647 12278 GCACCCACGGGCGGACAGAC 648 12279 CACCCACGGGCGGACAGACG 649 12280 ACCCACGGGCGGACAGACGG 650 12281 CCCACGGGCGGACAGACGGC 651 12282 CCACGGGCGGACAGACGGCC 652 12283 CACGGGCGGACAGACGGCCA 653 12284 CCGGCAGAATGGGCGCATTT 611 12285 GCCGGCAGAATGGGCGCATT 610 12286 AGCCGGCAGAATGGGCGCAT 609 12287 ACGCCACGAGGGCACCCACGGGCGGA 637 12288 CGCCACGAGGGCACCCACGG 638 12289 GCCACGAGGGCACCCACGGG 639 12290 CCACGAGGGCACCCACGGGC 640 12291 CACGAGGGCACCCACGGGCG 641 12292 ACGAGGGCACCCACGGGCGG 642 12293 CGAGGGCACCCACGGGCGGA 643 12294 GAGGGCACCCACGGGCGGAC 644 12295 AGGGCACCCACGGGCGGACA 645 12296 GGGCACCCACGGGCGGACAG 646 12297 GGCACCCACGGGCGGACAGA 647 12298 GCACCCACGGGCGGACAGAC 648 12299 CACCCACGGGCGGACAGACG 649 12300 ACCCACGGGCGGACAGACGG 650 12301 CCCACGGGCGGACAGACGGC 651 12302 CCACGGGCGGACAGACGGCC 652 12303 CACGGGCGGACAGACGGCCA 653 12304 AACGCCACGAGGGCACCCAC 636 12305 GAACGCCACGAGGGCACCCA 635 12306 AGAACGCCACGAGGGCACCC 634 12307 AAGAACGCCACGAGGGCACC 633 12308 CAAGAACGCCACGAGGGCAC 632 12309 CCAAGAACGCCACGAGGGCA 631 12310 TCCAAGAACGCCACGAGGGC 630 12311 TTCCAAGAACGCCACGAGGG 629 12312 TTTCCAAGAACGCCACGAGG 628 12313 ATTTCCAAGAACGCCACGAG 627 12314 CATTTCCAAGAACGCCACGA 626 12315 GCATTTCCAAGAACGCCACG 625 12316 CGCATTTCCAAGAACGCCAC 624 12317 GCGCATTTCCAAGAACGCCA 623 12318 GGCGCATTTCCAAGAACGCC 622 12319 GGGCGCATTTCCAAGAACGC 621 12320 TGGGCGCATTTCCAAGAACG 620 12321 ATGGGCGCATTTCCAAGAAC 619 12322 AATGGGCGCATTTCCAAGAA 618 12323 GAATGGGCGCATTTCCAAGA 617 12324 AGAATGGGCGCATTTCCAAG 616 12325 CAGAATGGGCGCATTTCCAA 615 12326 GCAGAATGGGCGCATTTCCA 614 12327 GGCAGAATGGGCGCATTTCC 613 12328 CGGCAGAATGGGCGCATTTC 612 12329 CCGGCAGAATGGGCGCATTT 611 12330 GCCGGCAGAATGGGCGCATT 610 12331 AGCCGGCAGAATGGGCGCAT 609 12332 CGGTGACCGCGGCCTGGGCGGATGG 2755 12333 GGTGACCGCGGCCTGGGCGG 2756 12334 GTGACCGCGGCCTGGGCGGA 2757 12335 TGACCGCGGCCTGGGCGGAT 2758 12336 GACCGCGGCCTGGGCGGATG 2759 12337 ACCGCGGCCTGGGCGGATGG 2760 12338 CCGCGGCCTGGGCGGATGGT 2761 12339 CGCGGCCTGGGCGGATGGTC 2762 12340 GCGGCCTGGGCGGATGGTCG 2763 12341 CGGCCTGGGCGGATGGTCGG 2764 12342 GGCCTGGGCGGATGGTCGGT 2765 12343 GCCTGGGCGGATGGTCGGTC 2766 12344 CCTGGGCGGATGGTCGGTCA 2767 12345 CTGGGCGGATGGTCGGTCAG 2768 12346 TGGGCGGATGGTCGGTCAGG 2769 12347 CCGGTGACCGCGGCCTGGGC 2754 12348 CCCGGTGACCGCGGCCTGGG 2753 12349 CCCCGGTGACCGCGGCCTGG 2752 12350 GCCCCGGTGACCGCGGCCTG 2751 12351 CGCCCCGGTGACCGCGGCCT 2750 12352 CCGCCCCGGTGACCGCGGCC 2749 12353 CCCGCCCCGGTGACCGCGGC 2748 12354 CCCCGCCCCGGTGACCGCGG 2747 12355 CCCCCGCCCCGGTGACCGCG 2746 12356 GCCCCCGCCCCGGTGACCGC 2745 12357 GGCCCCCGCCCCGGTGACCG 2744 12358 TGGCCCCCGCCCCGGTGACC 2743 12359 CTGGCCCCCGCCCCGGTGAC 2742 12360 CCTGGCCCCCGCCCCGGTGA 2741 12361 CCCTGGCCCCCGCCCCGGTG 2740 12362 CCCCTGGCCCCCGCCCCGGT 2739 12363 CCCCCTGGCCCCCGCCCCGG 2738 12364 GCCCCCTGGCCCCCGCCCCG 2737 12365 CGCCCCCTGGCCCCCGCCCC 2736 12366 TCGCCCCCTGGCCCCCGCCC 2735 12367 CTCGCCCCCTGGCCCCCGCC 2734 12368 CCTCGCCCCCTGGCCCCCGC 2733 12369 TCCTCGCCCCCTGGCCCCCG 2732 12370 TTCCTCGCCCCCTGGCCCCC 2731 12371 TTTCCTCGCCCCCTGGCCCC 2730 12372 CTTTCCTCGCCCCCTGGCCC 2729 12373 GCTTTCCTCGCCCCCTGGCC 2728 12374 CGCTTTCCTCGCCCCCTGGC 2727 12375 ACGCTTTCCTCGCCCCCTGG 2726 12376 CACGCTTTCCTCGCCCCCTG 2725 12377 TCACGCTTTCCTCGCCCCCT 2724 12378 TTCACGCTTTCCTCGCCCCC 2723 12379 CTTCACGCTTTCCTCGCCCC 2722 12380 CCTTCACGCTTTCCTCGCCC 2721 12381 ACCTTCACGCTTTCCTCGCC 2720 12382 CACCTTCACGCTTTCCTCGC 2719 12383 TCACCTTCACGCTTTCCTCG 2718 12384 ATCACCTTCACGCTTTCCTC 2717 12385 AATCACCTTCACGCTTTCCT 2716 12386 AAATCACCTTCACGCTTTCC 2715 12387 GAAATCACCTTCACGCTTTC 2714 12388 CGGGACTCAGCGCGGCTGCGCGCCG 2907 12389 GGGACTCAGCGCGGCTGCGC 2908 12390 GGACTCAGCGCGGCTGCGCG 2909 12391 GACTCAGCGCGGCTGCGCGC 2910 12392 ACTCAGCGCGGCTGCGCGCC 2911 12393 CTCAGCGCGGCTGCGCGCCG 2912 12394 TCAGCGCGGCTGCGCGCCGC 2913 12395 CAGCGCGGCTGCGCGCCGCG 2914 12396 AGCGCGGCTGCGCGCCGCGG 2915 12397 GCGCGGCTGCGCGCCGCGGG 2916 12398 CGCGGCTGCGCGCCGCGGGG 2917 12399 GCGGCTGCGCGCCGCGGGGC 2918 12400 CGGCTGCGCGCCGCGGGGCT 2919 12401 GGCTGCGCGCCGCGGGGCTC 2920 12402 GCTGCGCGCCGCGGGGCTCG 2921 12403 CTGCGCGCCGCGGGGCTCGG 2922 12404 TGCGCGCCGCGGGGCTCGGG 2923 12405 GCGCGCCGCGGGGCTCGGGG 2924 12406 CGCGCCGCGGGGCTCGGGGC 2925 12407 GCGCCGCGGGGCTCGGGGCT 2926 12408 CGCCGCGGGGCTCGGGGCTT 2927 12409 GCCGCGGGGCTCGGGGCTTG 2928 12410 CCGCGGGGCTCGGGGCTTGG 2929 12411 CGCGGGGCTCGGGGCTTGGG 2930 12412 GCGGGGCTCGGGGCTTGGGT 2931 12413 CGGGGCTCGGGGCTTGGGTT 2932 12414 GGGGCTCGGGGCTTGGGTTG 2933 12415 GGGCTCGGGGCTTGGGTTGG 2934 12416 GGCTCGGGGCTTGGGTTGGG 2935 12417 GCTCGGGGCTTGGGTTGGGG 2936 12418 CTCGGGGCTTGGGTTGGGGG 2937 12419 TCGGGGCTTGGGTTGGGGGC 2938 12420 CGGGGCTTGGGTTGGGGGCG 2939 12421 CCGGGACTCAGCGCGGCTGC 2906 12422 CCCGGGACTCAGCGCGGCTG 2905 12423 CCCCGGGACTCAGCGCGGCT 2904 12424 ACCCCGGGACTCAGCGCGGC 2903 12425 GACCCCGGGACTCAGCGCGG 2902 12426 AGACCCCGGGACTCAGCGCG 2901 12427 CAGACCCCGGGACTCAGCGC 2900 12428 GCAGACCCCGGGACTCAGCG 2899 12429 CGCAGACCCCGGGACTCAGC 2898 12430 ACGCAGACCCCGGGACTCAG 2897 12431 GACGCAGACCCCGGGACTCA 2896 12432 CGACGCAGACCCCGGGACTC 2895 12433 GCGACGCAGACCCCGGGACT 2894 12434 CGCGACGCAGACCCCGGGAC 2893 12435 CCGCGACGCAGACCCCGGGA 2892 12436 GCCGCGACGCAGACCCCGGG 2891 12437 CGCCGCGACGCAGACCCCGG 2890 12438 GCGCCGCGACGCAGACCCCG 2889 12439 CGCGCCGCGACGCAGACCCC 2888 12440 GCGCGCCGCGACGCAGACCC 2887 12441 GGCGCGCCGCGACGCAGACC 2886 12442 CGGCGCGCCGCGACGCAGAC 2885 12443 CCGGCGCGCCGCGACGCAGA 2884 12444 ACCGGCGCGCCGCGACGCAG 2883 12445 AACCGGCGCGCCGCGACGCA 2882 12446 GAACCGGCGCGCCGCGACGC 2881 12447 GGAACCGGCGCGCCGCGACG 2880 12448 AGGAACCGGCGCGCCGCGAC 2879 12449 CAGGAACCGGCGCGCCGCGA 2878 12450 TCAGGAACCGGCGCGCCGCG 2877 12451 TTCAGGAACCGGCGCGCCGC 2876 12452 ATTCAGGAACCGGCGCGCCG 2875 12453 CATTCAGGAACCGGCGCGCC 2874 12454 TCATTCAGGAACCGGCGCGC 2873 12455 TTCATTCAGGAACCGGCGCG 2872 12456 GTTCATTCAGGAACCGGCGC 2871 12457 CGTTCATTCAGGAACCGGCG 2870 12458 GCGTTCATTCAGGAACCGGC 2869 12459 CGCGTTCATTCAGGAACCGG 2868 12460 GCGCGTTCATTCAGGAACCG 2867 12461 AGCGCGTTCATTCAGGAACC 2866 12462 GAGCGCGTTCATTCAGGAAC 2865 12463 GGAGCGCGTTCATTCAGGAA 2864 12464 GGGAGCGCGTTCATTCAGGA 2863 12465 AGGGAGCGCGTTCATTCAGG 2862 12466 AAGGGAGCGCGTTCATTCAG 2861 12467 GAAGGGAGCGCGTTCATTCA 2860 12468 GGAAGGGAGCGCGTTCATTC 2859 12469 GGGAAGGGAGCGCGTTCATT 2858

Hot Zones (Relative upstream location to gene start site)  1-250 550-700 2700-2300

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13676) GGCCCTGCCGCCCAGAACTCGCTGGGCAAGTCGTGCCCCGCGTGAACACA CAGAAGGGGCTTGGGGACCGAGCGCGGCCCATCAGTCCCTCAGACCCTGA GGACCCAGAATTCCCTAAGGGGTCCGAATCCGAGTCCTGCCCCCAGCCCT TAAGGCACGGGCTCCAGGGACCCCAGGGGAAGGGCGCGGGGCATTAGGTA CGCAACCCGTTTCCCCGCACCTGGAAAAAAACTCCCTTTCCCTCCCCTCC CCTGCTTGTTGAGTGTCCGGATAACCAGAACTCTAAGGCGCCCCGTAATA ACGACCCCGCTGTCCCTCCACCCACCCCCAAGTGCCAAAGCGAGGGATGG AAGCGCTTTCAAGCGTTCCAAGGGCATTGAGGAGCGAGCTGGAGAGGCGC GGGGATGCGGGGTCCTCCCCGCAGTCTTCCGGAAAGGGCGGGGGAGGGCG CGGCAAGTTCCGGAGTGGGGCATGCCGTGGGAGCCCACGAGGGCCTCAGC GCGGATCCTCCGCCGGAAAACCGGCTCCCGCGAGCCGCCGCCGCAGGTTT CCTAGGCCCCGCGAGTCCCGCAGCGAAGCCCTGCGTCTCCGTCCGACGCG GGGGTCTGCTCAGCCTCGGGTGGGCCGCGGCCAGGCCTGACTGCGGGGGA GAGGGCCGAACGTGACCTCCGAGGTCACCCCCAGCCAGCTTTCTCTCCTG TGGTCGGAAGTGGTTTTCTTCTCGATCTGGGCGCCTACTCCCCACCACTT GGTCTGAGAGGGGCTGGGGCCGGAAGGCCAGGGAATCTCTGGTGGATTTG GGGGTTCATATTGCTCAGGGTACCAGCCGATGCGTTTTGAGGGGCGGGAG TCGAGGAATTAGAATCGCCTTTAACCCTCAAGAGTTGCGCCTTCAGCCTC GGGATCCCAGATGCGTCGTTGGAGCCAGGGCCGCCCCCCTACCTGTTGGG TTTGCGTTTTAACTCCAGCGCACACCTTGCCGGCAGCCCTCGGAGCTAGG GGAGGGGTCTCGTTTCCCCGCAGCCCGCCGGACAGACGACTGGGGCACGG GAGGGGCGGTGGCAGGGTGGTCTGTGTGTGGCTGAAACTAATTGATCTGG AGCGGAAACGCACGTCTGCGGTTGGGGCGATGGGGGGGGCGGTGCGGCTG TCCATGTGCCGAGCGTGTGGCTGTCTCGGGTGGGCACTGGGGCCGGAGTT CGCCCCGGCCCACCTCGCAGTTTTGGGGCGCCTGGGATCGGCGCTACGTA AGCGAAGCAGAGCTGCCATAGCACGTGGGCCGCCACGCGCACCCCAAAAG CAAGCAGTGTGGGGGGAAGGGGAGCTCGAGCGCCTTCGGAGCCCAGGGGC CGGCTTTCGGAAGCGTTTTCCCGGGCGACTTAAGGGCTTAACAATGGAAA ACTCGCGGAGCCTGAGCCAAGTCCTTTCAAGTCGCCGCCAGGTATGCGGC TGCAGGTGACCCCACCTGGGTGCGCCCGCCCGCCAGCCGCCCTGGTGGAA AAGCGGGTGCGGGAGGTCGCTGGCGAAAGGTCGGGACTGGTCCCTGCACC ACCCGCCCCCAACCCAAGCCCCGAGCCCCGCGGCGCGCAGCCGCGCTGAG TCCCGGGGTCTGCGTCGCGGCGCGCCGGTTCCTGAATGAACGCGCTCCCT TCCCCCGCCTGAATGAAGGTTCCCACAGCCAGGGACGGTGGCGAACACGC GCCTGCAGCGGAATTCGCTTTCTCCTGACCGACCATCCGCCCAGGCCGCG GTCACCGGGGCGGGGGCCAGGGGGCGAGGAAAGCGTGAAGGTGATTTCAG TTAATTTTGGATTTTCTTTCAAACAACGTGGTTACCCTCCCGACTGGGCC ACTTGCCCTTTGTCTCCAAATGGTCACCAAGAAATAAGAACAGAGCACTT TAAATGAGCCCAGAATCCGCAGTTCCTGCTTCGTGGTGGGTTTTAAGAAG ACAGTGTAAAGTAAAACTGCAACCGAAAAGTTTTTTAAAGTTGCTTTTCT CTTTGGAAAAAATAAAATCAAAATGCTTTCTCTGCGCTTCTTGAAGCAAT GACCCTCAAAAGCCCAGAGGTATTGGCCCCCTCGGGGGACCCGGGGGCCG CCAAGCAGGGTTCCCCCAGGTGGGGGCTGGGCAGCTGGCGCTCCCCGCCG GGCCCCAAATTCCAGCGCCGGGCCCCAAATTCCAGCGCCTCCCCCGCGGG TTCCTGGACGGCTCTTTACGCTCGCTAACCGGGCTTGCAATTTTGCGCTC GTCCCTGAGCCGGGAAATCAACGAAGTTCCTAGTCGAGATCTGCCCGGTC CGCCTAGTAACAGCGCCGCGCCCCCATTGGCTCATGCTAATTCCAGTTTC CTCTGTCTTGCGCCCGGGATGGGGGGGTGAAGCTCCCTCCTGGACCCAGA GCCGGTTGTGCCGGAGTGGGCGAGCCTCTTTATGCCCTGCTGCCCCTAGC CGACTTCGGCCCGCTTCGCGCCTCGGGCTGGGCCAGGGCGCACGCGGGGC TCGGGGCCCCTCGCCCCACGGGATGGGAGAGGCCGGGTGATAGCTCCGGG CCCCATAAATCATCCAGGCGGCCGCCGGGTCGGGATTTTATGAATGAAAA AGCAGCTGGGCCGCCCTTGTGCGCGGGCTGATGCTCTGAGGCTTGGCTAT GCGGGGGCCAACGCGATTGTGGGTGCTCGGGGAGTGGGGGGGGGCACGAC CGTAGGTGCTCCCTGCTGGGGCAACCCATCGCTCCCCATGCGGAATCCGG GGGTAATTACCCCCCCAGGACCCGGAATATTAGTAATCCTAATTCCCGGC GGGGGAGGGGGCGCGGGAGGAATTCACCCTGAAAGGTGGGGGTGGGGGGG GTCGCATCTTGCTGTGAGCACCCTGGCGAAGGGGAGAGGGCTTTTTCTAT CAGTTTTCTTTGAGCTTTTACTGTTAAGAGGGTACGGTGGTTTGATGACA CTGAACTATATTCAAAAGGAAGTAAATGAACAGTTTTCTTAATTTGGGGC AGGTACTGTAAAAATAAAAACAAAAGTTAAGACAGTAAAATGTCCTTTTA TTTTTTAATGCACCAAAGAGACAGAACCTGTAATTTTAAAAACTGTGTAT TTTAATTTACATCTGCTTAAGTTTGCGATAATATTGGGGACCCTCTCATG TAACCACGAACACCTATCGATTTTGCTAAAAATCAGATCAGTACACTCGT TTGTTTAATTGATAATTGTTCTGAATTATGCCGGCTCCTGCCAGCCCCCT CACGCTCACGAATTCAGTCCCAGGGCAAATTCTAAAGGTGAAGGGACGTC TACACCCCCAACAAAACCAATTAGGAACCTTCGGTGGTCTTGTCCCAGGC AGAGGGGACTAATATTTCCAGCAATTTAATTTCTTTTTTAATTAAAAAAA ATGAGTCAGAATGGAGATCACTGTTTCTCAGCTTTCCATTCAGAGGTGTG TTTCTCCCGGTTAAATTGCCGGCACGGGAAGGGAGGGGGTGCAGTTGGGG ACCCCCGCAAGGACCGACTGGTCAAGGTAGGAAGGCAGCCCGAAGAGTCT CCAGGCTAGAAGGACAAGATGAAGGAAATGCTGGCCACCATCTTGGGCTG CTGCTGGAATTTTCGGGCATTTATTTTATTTTATTTTTTGAGCGAGCGCA TGCTAAGCTGAAATCCCTTTAACTTTTAGGGTTACCCCCTTGGGCATTTG CAACGACGCCCCTGTGCGCCGGAATGAAACTTGCACAGGGGTTGTGTGCC CGGTCCTCCCCGTCCTTGCATGCTAAATTAGTTCTTGCAATTTACACGTG TTAATGAAAATGAAAGAAGATGCAGTCGCTGAGATTCTTTGGCCGTCTGT CCGCCCGTGGGTGCCCTCGTGGCGTTCTTGGAAATGCGCCCATTCTGCCG GCTTGGATATGGGGTGTCGCCGCGCCCCAGTCACCCCTTCTCGTGGTCTC CCCAGGCTGCGTGTGGCCTGCCGGCCTTCCTAGTTGTCCCCTACTGCAGA GCCACCTCCACCTCACCCCCTAAATCCCGGGGGACCCACTCGAGGCGGAC GGGGCCCCCTGCACCCCTCTTCCCTGGCGGGGAGAAAGGCTGCAGCGGGG CGATTTGCATTTCTATGAAAACCGGACTACAGGGGCAACTCCGCCGCAGG GCAGGCGCGGCGCCTCAGGGATGGCTTTTGGGCTCTGCCCCTCGCTGCTC CCGGCGTTTGGCGCCCGCGCCCCCTCCCCCTGCGCCCGCCCCCGCCCCCC TCCCGCTCCCATTCTCTGCCGGGCTTTGATCTTTGCTTAACAACAGTAAC GTCACACGGACTACAGGGGAGTTTTGTTGAAGTTGCAAAGTCCTGGAGCC TCCAGAGGGCTGTCGGCGCAGTAGCAGCGAGCAGCAGAGTCCGCACGCTC CGGCGAGGGGCAGAAGAGCGCGAGGGAGCGCGGGGCAGCAGAAGCGAGAG CCGAGCGCGGACCCAGCCAGGACCCACAGCCCTCCCCAGCTGCCCAGGAA GAGCCCCAGCC ATG

MIF1

MIF1 (macrophage migration inhibitory factor 1) is a lymphokine involved in cell-mediated immunity, immunoregulation, and inflammation. MIF forms a homotrimer with three catalytic sites. The MIF homotrimer can enter a cell via endocytosis where it interacts with intracellular proteins. This interaction results in downregulating MAPK signals leading to activation of Cyclin D1 and subsequent cellular proliferation. Depending on the cellular environment, MIF may also have antioxidant activity which would inhibit apoptosis. Apoptosis can also be inhibited via a MIF-CD74 complex. MIF has been associated with inflammation, including rheumatoid arthritis, sepsis, and cancer.

Protein: MIF1 Gene: MIF1 (Homo sapiens, chromosome 22, 24236565-24237409[NCBI Reference Sequence: NC_(—)000022.10]; start site location: 24236662; strand: positive)

Gene Identification GeneID 4282 HGNC 7097 HPRD 01091 MIM 153620

Targeted Sequences Relative upstream Sequence Design location to gene start ID No: ID Sequence (5′-3′) site 12470 GACCCGCGCAGAGGCACAGACGC 22 12490 CGCCACCGCCGGCGCCAGGCCCCGCCCCCGCG 123 12701 CGTTCCTCCAGCAACCGCCGCTAAGCCCGGCG 238 12902 CGCCTGCCTCGGCTCGACCCCCGCAG 182 13123 CGGCTAGAAATCGGCCTGTTCCGGCCTCGCCT 297 13174 CGGGGGTGGGGATGCGGCGGTGAACCCG 384 13175 CGCGGCAGGTGAGAGGGGAGCTGCCCCTGCG 568 13176 CGCGTGCACGTGTGTCCACATGAGTGC 3656 13203 MIF1_1 CGCCACCGCCGGCGCCAGGCCCCGCC 117 13414 MIF1_2 CGCGGCAGGTGAGAGGGGAGCTGCCC 563

Target Shift Sequences Relative upstream location to gene Sequence start ID No: Sequence (5′-3′) site 12470 GACCCGCGCAGAGGCACAGACGC 20 12471 ACCCGCGCAGAGGCACAGAC 21 12472 CCCGCGCAGAGGCACAGACG 22 12473 CCGCGCAGAGGCACAGACGC 23 12474 CGCGCAGAGGCACAGACGCA 24 12475 GCGCAGAGGCACAGACGCAC 25 12476 CGCAGAGGCACAGACGCACG 26 12477 GCAGAGGCACAGACGCACGC 27 12478 CAGAGGCACAGACGCACGCG 28 12479 AGAGGCACAGACGCACGCGC 29 12480 GAGGCACAGACGCACGCGCC 30 12481 AGGCACAGACGCACGCGCCG 31 12482 GGCACAGACGCACGCGCCGC 32 12483 GCACAGACGCACGCGCCGCG 33 12484 CACAGACGCACGCGCCGCGG 34 12485 ACAGACGCACGCGCCGCGGC 35 12486 CAGACGCACGCGCCGCGGCC 36 12487 AGACCCGCGCAGAGGCACAG 19 12488 GAGACCCGCGCAGAGGCACA 18 12489 GGAGACCCGCGCAGAGGCAC 17 12490 CGCCACCGCCGGCGCCAGGC 112 CCCGCCCCCGCG 12491 GCCACCGCCGGCGCCAGGCC 113 12492 CCACCGCCGGCGCCAGGCCC 114 12493 CACCGCCGGCGCCAGGCCCC 115 12494 ACCGCCGGCGCCAGGCCCCG 116 12495 CCGCCGGCGCCAGGCCCCGC 117 12496 CGCCGGCGCCAGGCCCCGCC 118 12497 GCCGGCGCCAGGCCCCGCCC 119 12498 CCGGCGCCAGGCCCCGCCCC 120 12499 CGGCGCCAGGCCCCGCCCCC 121 12500 GGCGCCAGGCCCCGCCCCCG 122 12501 GCGCCAGGCCCCGCCCCCGC 123 12502 CGCCAGGCCCCGCCCCCGCG 124 12503 GCCAGGCCCCGCCCCCGCGA 125 12504 CCAGGCCCCGCCCCCGCGAG 126 12505 CAGGCCCCGCCCCCGCGAGG 127 12506 AGGCCCCGCCCCCGCGAGGC 128 12507 GGCCCCGCCCCCGCGAGGCT 129 12508 GCCCCGCCCCCGCGAGGCTG 130 12509 CCCCGCCCCCGCGAGGCTGC 131 12510 CCCGCCCCCGCGAGGCTGCG 132 12511 CCGCCCCCGCGAGGCTGCGG 133 12512 CGCCCCCGCGAGGCTGCGGC 134 12513 GCCCCCGCGAGGCTGCGGCT 135 12514 CCCCCGCGAGGCTGCGGCTC 136 12515 CCCCGCGAGGCTGCGGCTCC 137 12516 CCCGCGAGGCTGCGGCTCCG 138 12517 CCGCGAGGCTGCGGCTCCGC 139 12518 CGCGAGGCTGCGGCTCCGCC 140 12519 GCGAGGCTGCGGCTCCGCCC 141 12520 CGAGGCTGCGGCTCCGCCCC 142 12521 GAGGCTGCGGCTCCGCCCCG 143 12522 AGGCTGCGGCTCCGCCCCGA 144 12523 GGCTGCGGCTCCGCCCCGAG 145 12524 GCTGCGGCTCCGCCCCGAGT 146 12525 CTGCGGCTCCGCCCCGAGTG 147 12526 TGCGGCTCCGCCCCGAGTGG 148 12527 GCGGCTCCGCCCCGAGTGGG 149 12528 CGGCTCCGCCCCGAGTGGGG 150 12529 GGCTCCGCCCCGAGTGGGGA 151 12530 GCTCCGCCCCGAGTGGGGAA 152 12531 CTCCGCCCCGAGTGGGGAAG 153 12532 TCCGCCCCGAGTGGGGAAGT 154 12533 CCGCCCCGAGTGGGGAAGTC 155 12534 CGCCCCGAGTGGGGAAGTCA 156 12535 GCCCCGAGTGGGGAAGTCAC 157 12536 CCCCGAGTGGGGAAGTCACC 158 12537 CCCGAGTGGGGAAGTCACCG 159 12538 CCGAGTGGGGAAGTCACCGC 160 12539 CGAGTGGGGAAGTCACCGCC 161 12540 GAGTGGGGAAGTCACCGCCT 162 12541 AGTGGGGAAGTCACCGCCTG 163 12542 GTGGGGAAGTCACCGCCTGC 164 12543 TGGGGAAGTCACCGCCTGCC 165 12544 GGGGAAGTCACCGCCTGCCT 166 12545 GGGAAGTCACCGCCTGCCTC 167 12546 GGAAGTCACCGCCTGCCTCG 168 12547 GAAGTCACCGCCTGCCTCGG 169 12548 AAGTCACCGCCTGCCTCGGC 170 12549 AGTCACCGCCTGCCTCGGCT 171 12550 GTCACCGCCTGCCTCGGCTC 172 12551 TCACCGCCTGCCTCGGCTCG 173 12552 CACCGCCTGCCTCGGCTCGA 174 12553 ACCGCCTGCCTCGGCTCGAC 175 12554 CCGCCTGCCTCGGCTCGACC 176 12555 CGCCTGCCTCGGCTCGACCC 177 12556 GCCTGCCTCGGCTCGACCCC 178 12557 CCTGCCTCGGCTCGACCCCC 179 12558 CTGCCTCGGCTCGACCCCCG 180 12559 TGCCTCGGCTCGACCCCCGC 181 12560 GCCTCGGCTCGACCCCCGCA 182 12561 CCTCGGCTCGACCCCCGCAG 183 12562 CTCGGCTCGACCCCCGCAGG 184 12563 TCGGCTCGACCCCCGCAGGG 185 12564 CGGCTCGACCCCCGCAGGGC 186 12565 GGCTCGACCCCCGCAGGGCA 187 12566 GCTCGACCCCCGCAGGGCAG 188 12567 CTCGACCCCCGCAGGGCAGG 189 12568 TCGACCCCCGCAGGGCAGGA 190 12569 CGACCCCCGCAGGGCAGGAC 191 12570 GACCCCCGCAGGGCAGGACC 192 12571 ACCCCCGCAGGGCAGGACCC 193 12572 CCCCCGCAGGGCAGGACCCT 194 12573 CCCCGCAGGGCAGGACCCTG 195 12574 CCCGCAGGGCAGGACCCTGG 196 12575 CCGCAGGGCAGGACCCTGGG 197 12576 CGCAGGGCAGGACCCTGGGC 198 12577 GCAGGGCAGGACCCTGGGCG 199 12578 CAGGGCAGGACCCTGGGCGA 200 12579 AGGGCAGGACCCTGGGCGAC 201 12580 GGGCAGGACCCTGGGCGACT 202 12581 GGCAGGACCCTGGGCGACTC 203 12582 GCAGGACCCTGGGCGACTCC 204 12583 CAGGACCCTGGGCGACTCCG 205 12584 AGGACCCTGGGCGACTCCGC 206 12585 GGACCCTGGGCGACTCCGCC 207 12586 GACCCTGGGCGACTCCGCCC 208 12587 ACCCTGGGCGACTCCGCCCG 209 12588 CCCTGGGCGACTCCGCCCGT 210 12589 CCTGGGCGACTCCGCCCGTT 211 12590 CTGGGCGACTCCGCCCGTTC 212 12591 TGGGCGACTCCGCCCGTTCC 213 12592 GGGCGACTCCGCCCGTTCCT 214 12593 GGCGACTCCGCCCGTTCCTC 215 12594 GCGACTCCGCCCGTTCCTCC 216 12595 CGACTCCGCCCGTTCCTCCA 217 12596 GACTCCGCCCGTTCCTCCAG 218 12597 ACTCCGCCCGTTCCTCCAGC 219 12598 CTCCGCCCGTTCCTCCAGCA 220 12599 TCCGCCCGTTCCTCCAGCAA 221 12600 CCGCCCGTTCCTCCAGCAAC 222 12601 CGCCCGTTCCTCCAGCAACC 223 12602 GCCCGTTCCTCCAGCAACCG 224 12603 CCCGTTCCTCCAGCAACCGC 225 12604 CCGTTCCTCCAGCAACCGCC 226 12605 CGTTCCTCCAGCAACCGCCG 227 12606 GTTCCTCCAGCAACCGCCGC 228 12607 TTCCTCCAGCAACCGCCGCT 229 12608 TCCTCCAGCAACCGCCGCTA 230 12609 CCTCCAGCAACCGCCGCTAA 231 12610 CTCCAGCAACCGCCGCTAAG 232 12611 TCCAGCAACCGCCGCTAAGC 233 12612 CCAGCAACCGCCGCTAAGCC 234 12613 CAGCAACCGCCGCTAAGCCC 235 12614 AGCAACCGCCGCTAAGCCCG 236 12615 GCAACCGCCGCTAAGCCCGG 237 12616 CAACCGCCGCTAAGCCCGGC 238 12617 AACCGCCGCTAAGCCCGGCG 239 12618 ACCGCCGCTAAGCCCGGCGC 240 12619 CCGCCGCTAAGCCCGGCGCA 241 12620 CGCCGCTAAGCCCGGCGCAC 242 12621 GCCGCTAAGCCCGGCGCACC 243 12622 CCGCTAAGCCCGGCGCACCG 244 12623 CGCTAAGCCCGGCGCACCGC 245 12624 GCTAAGCCCGGCGCACCGCT 246 12625 CTAAGCCCGGCGCACCGCTC 247 12626 TAAGCCCGGCGCACCGCTCC 248 12627 AAGCCCGGCGCACCGCTCCA 249 12628 AGCCCGGCGCACCGCTCCAA 250 12629 GCCCGGCGCACCGCTCCAAC 251 12630 CCCGGCGCACCGCTCCAACC 252 12631 CCGGCGCACCGCTCCAACCT 253 12632 CGGCGCACCGCTCCAACCTG 254 12633 GGCGCACCGCTCCAACCTGT 255 12634 GCGCACCGCTCCAACCTGTT 256 12635 CGCACCGCTCCAACCTGTTC 257 12636 GCACCGCTCCAACCTGTTCT 258 12637 CACCGCTCCAACCTGTTCTC 259 12638 ACCGCTCCAACCTGTTCTCC 260 12639 CCGCTCCAACCTGTTCTCCA 261 12640 CGCTCCAACCTGTTCTCCAC 262 12641 ACGCCACCGCCGGCGCCAGG 111 12642 GACGCCACCGCCGGCGCCAG 110 12643 TGACGCCACCGCCGGCGCCA 109 12644 GTGACGCCACCGCCGGCGCC 108 12645 TGTGACGCCACCGCCGGCGC 107 12646 TTGTGACGCCACCGCCGGCG 106 12647 TTTGTGACGCCACCGCCGGC 105 12648 TTTTGTGACGCCACCGCCGG 104 12649 CTTTTGTGACGCCACCGCCG 103 12650 CCTTTTGTGACGCCACCGCC 102 12651 GCCTTTTGTGACGCCACCGC 101 12652 CGCCTTTTGTGACGCCACCG 100 12653 CCGCCTTTTGTGACGCCACC 99 12654 CCCGCCTTTTGTGACGCCAC 98 12655 TCCCGCCTTTTGTGACGCCA 97 12656 GTCCCGCCTTTTGTGACGCC 96 12657 GGTCCCGCCTTTTGTGACGC 95 12658 TGGTCCCGCCTTTTGTGACG 94 12659 GTGGTCCCGCCTTTTGTGAC 93 12660 TGTGGTCCCGCCTTTTGTGA 92 12661 CTGTGGTCCCGCCTTTTGTG 91 12662 ACTGTGGTCCCGCCTTTTGT 90 12663 CACTGTGGTCCCGCCTTTTG 89 12664 CCACTGTGGTCCCGCCTTTT 88 12665 ACCACTGTGGTCCCGCCTTT 87 12666 CACCACTGTGGTCCCGCCTT 86 12667 ACACCACTGTGGTCCCGCCT 85 12668 GACACCACTGTGGTCCCGCC 84 12669 GGACACCACTGTGGTCCCGC 83 12670 CGGACACCACTGTGGTCCCG 82 12671 TCGGACACCACTGTGGTCCC 81 12672 CTCGGACACCACTGTGGTCC 80 12673 TCTCGGACACCACTGTGGTC 79 12674 TTCTCGGACACCACTGTGGT 78 12675 CTTCTCGGACACCACTGTGG 77 12676 ACTTCTCGGACACCACTGTG 76 12677 GACTTCTCGGACACCACTGT 75 12678 TGACTTCTCGGACACCACTG 74 12679 CTGACTTCTCGGACACCACT 73 12680 CCTGACTTCTCGGACACCAC 72 12681 GCCTGACTTCTCGGACACCA 71 12682 TGCCTGACTTCTCGGACACC 70 12683 GTGCCTGACTTCTCGGACAC 69 12684 CGTGCCTGACTTCTCGGACA 68 12685 ACGTGCCTGACTTCTCGGAC 67 12686 TACGTGCCTGACTTCTCGGA 66 12687 CTACGTGCCTGACTTCTCGG 65 12688 GCTACGTGCCTGACTTCTCG 64 12689 AGCTACGTGCCTGACTTCTC 63 12690 GAGCTACGTGCCTGACTTCT 62 12691 TGAGCTACGTGCCTGACTTC 61 12692 CTGAGCTACGTGCCTGACTT 60 12693 GCTGAGCTACGTGCCTGACT 59 12694 CGCTGAGCTACGTGCCTGAC 58 12695 CCGCTGAGCTACGTGCCTGA 57 12696 GCCGCTGAGCTACGTGCCTG 56 12697 CGCCGCTGAGCTACGTGCCT 55 12698 CCGCCGCTGAGCTACGTGCC 54 12699 GCCGCCGCTGAGCTACGTGC 53 12700 GGCCGCCGCTGAGCTACGTG 52 12701 CGTTCCTCCAGCAACCGCCGCTAAGCCCGGCG 227 12702 GTTCCTCCAGCAACCGCCGC 228 12703 TTCCTCCAGCAACCGCCGCT 229 12704 TCCTCCAGCAACCGCCGCTA 230 12705 CCTCCAGCAACCGCCGCTAA 231 12706 CTCCAGCAACCGCCGCTAAG 232 12707 TCCAGCAACCGCCGCTAAGC 233 12708 CCAGCAACCGCCGCTAAGCC 234 12709 CAGCAACCGCCGCTAAGCCC 235 12710 AGCAACCGCCGCTAAGCCCG 236 12711 GCAACCGCCGCTAAGCCCGG 237 12712 CAACCGCCGCTAAGCCCGGC 238 12713 AACCGCCGCTAAGCCCGGCG 239 12714 ACCGCCGCTAAGCCCGGCGC 240 12715 CCGCCGCTAAGCCCGGCGCA 241 12716 CGCCGCTAAGCCCGGCGCAC 242 12717 GCCGCTAAGCCCGGCGCACC 243 12718 CCGCTAAGCCCGGCGCACCG 244 12719 CGCTAAGCCCGGCGCACCGC 245 12720 GCTAAGCCCGGCGCACCGCT 246 12721 CTAAGCCCGGCGCACCGCTC 247 12722 TAAGCCCGGCGCACCGCTCC 248 12723 AAGCCCGGCGCACCGCTCCA 249 12724 AGCCCGGCGCACCGCTCCAA 250 12725 GCCCGGCGCACCGCTCCAAC 251 12726 CCCGGCGCACCGCTCCAACC 252 12727 CCGGCGCACCGCTCCAACCT 253 12728 CGGCGCACCGCTCCAACCTG 254 12729 GGCGCACCGCTCCAACCTGT 255 12730 GCGCACCGCTCCAACCTGTT 256 12731 CGCACCGCTCCAACCTGTTC 257 12732 GCACCGCTCCAACCTGTTCT 258 12733 CACCGCTCCAACCTGTTCTC 259 12734 ACCGCTCCAACCTGTTCTCC 260 12735 CCGCTCCAACCTGTTCTCCA 261 12736 CGCTCCAACCTGTTCTCCAC 262 12737 CCGTTCCTCCAGCAACCGCC 226 12738 CCCGTTCCTCCAGCAACCGC 225 12739 GCCCGTTCCTCCAGCAACCG 224 12740 CGCCCGTTCCTCCAGCAACC 223 12741 CCGCCCGTTCCTCCAGCAAC 222 12742 TCCGCCCGTTCCTCCAGCAA 221 12743 CTCCGCCCGTTCCTCCAGCA 220 12744 ACTCCGCCCGTTCCTCCAGC 219 12745 GACTCCGCCCGTTCCTCCAG 218 12746 CGACTCCGCCCGTTCCTCCA 217 12747 GCGACTCCGCCCGTTCCTCC 216 12748 GGCGACTCCGCCCGTTCCTC 215 12749 GGGCGACTCCGCCCGTTCCT 214 12750 TGGGCGACTCCGCCCGTTCC 213 12751 CTGGGCGACTCCGCCCGTTC 212 12752 CCTGGGCGACTCCGCCCGTT 211 12753 CCCTGGGCGACTCCGCCCGT 210 12754 ACCCTGGGCGACTCCGCCCG 209 12755 GACCCTGGGCGACTCCGCCC 208 12756 GGACCCTGGGCGACTCCGCC 207 12757 AGGACCCTGGGCGACTCCGC 206 12758 CAGGACCCTGGGCGACTCCG 205 12759 GCAGGACCCTGGGCGACTCC 204 12760 GGCAGGACCCTGGGCGACTC 203 12761 GGGCAGGACCCTGGGCGACT 202 12762 AGGGCAGGACCCTGGGCGAC 201 12763 CAGGGCAGGACCCTGGGCGA 200 12764 GCAGGGCAGGACCCTGGGCG 199 12765 CGCAGGGCAGGACCCTGGGC 198 12766 CCGCAGGGCAGGACCCTGGG 197 12767 CCCGCAGGGCAGGACCCTGG 196 12768 CCCCGCAGGGCAGGACCCTG 195 12769 CCCCCGCAGGGCAGGACCCT 194 12770 ACCCCCGCAGGGCAGGACCC 193 12771 GACCCCCGCAGGGCAGGACC 192 12772 CGACCCCCGCAGGGCAGGAC 191 12773 TCGACCCCCGCAGGGCAGGA 190 12774 CTCGACCCCCGCAGGGCAGG 189 12775 GCTCGACCCCCGCAGGGCAG 188 12776 GGCTCGACCCCCGCAGGGCA 187 12777 CGGCTCGACCCCCGCAGGGC 186 12778 TCGGCTCGACCCCCGCAGGG 185 12779 CTCGGCTCGACCCCCGCAGG 184 12780 CCTCGGCTCGACCCCCGCAG 183 12781 GCCTCGGCTCGACCCCCGCA 182 12782 TGCCTCGGCTCGACCCCCGC 181 12783 CTGCCTCGGCTCGACCCCCG 180 12784 CCTGCCTCGGCTCGACCCCC 179 12785 GCCTGCCTCGGCTCGACCCC 178 12786 CGCCTGCCTCGGCTCGACCC 177 12787 CCGCCTGCCTCGGCTCGACC 176 12788 ACCGCCTGCCTCGGCTCGAC 175 12789 CACCGCCTGCCTCGGCTCGA 174 12790 TCACCGCCTGCCTCGGCTCG 173 12791 GTCACCGCCTGCCTCGGCTC 172 12792 AGTCACCGCCTGCCTCGGCT 171 12793 AAGTCACCGCCTGCCTCGGC 170 12794 GAAGTCACCGCCTGCCTCGG 169 12795 GGAAGTCACCGCCTGCCTCG 168 12796 GGGAAGTCACCGCCTGCCTC 167 12797 GGGGAAGTCACCGCCTGCCT 166 12798 TGGGGAAGTCACCGCCTGCC 165 12799 GTGGGGAAGTCACCGCCTGC 164 12800 AGTGGGGAAGTCACCGCCTG 163 12801 GAGTGGGGAAGTCACCGCCT 162 12802 CGAGTGGGGAAGTCACCGCC 161 12803 CCGAGTGGGGAAGTCACCGC 160 12804 CCCGAGTGGGGAAGTCACCG 159 12805 CCCCGAGTGGGGAAGTCACC 158 12806 GCCCCGAGTGGGGAAGTCAC 157 12807 CGCCCCGAGTGGGGAAGTCA 156 12808 CCGCCCCGAGTGGGGAAGTC 155 12809 TCCGCCCCGAGTGGGGAAGT 154 12810 CTCCGCCCCGAGTGGGGAAG 153 12811 GCTCCGCCCCGAGTGGGGAA 152 12812 GGCTCCGCCCCGAGTGGGGA 151 12813 CGGCTCCGCCCCGAGTGGGG 150 12814 GCGGCTCCGCCCCGAGTGGG 149 12815 TGCGGCTCCGCCCCGAGTGG 148 12816 CTGCGGCTCCGCCCCGAGTG 147 12817 GCTGCGGCTCCGCCCCGAGT 146 12818 GGCTGCGGCTCCGCCCCGAG 145 12819 AGGCTGCGGCTCCGCCCCGA 144 12820 GAGGCTGCGGCTCCGCCCCG 143 12821 CGAGGCTGCGGCTCCGCCCC 142 12822 GCGAGGCTGCGGCTCCGCCC 141 12823 CGCGAGGCTGCGGCTCCGCC 140 12824 CCGCGAGGCTGCGGCTCCGC 139 12825 CCCGCGAGGCTGCGGCTCCG 138 12826 CCCCGCGAGGCTGCGGCTCC 137 12827 CCCCCGCGAGGCTGCGGCTC 136 12828 GCCCCCGCGAGGCTGCGGCT 135 12829 CGCCCCCGCGAGGCTGCGGC 134 12830 CCGCCCCCGCGAGGCTGCGG 133 12831 CCCGCCCCCGCGAGGCTGCG 132 12832 CCCCGCCCCCGCGAGGCTGC 131 12833 GCCCCGCCCCCGCGAGGCTG 130 12834 GGCCCCGCCCCCGCGAGGCT 129 12835 AGGCCCCGCCCCCGCGAGGC 128 12836 CAGGCCCCGCCCCCGCGAGG 127 12837 CCAGGCCCCGCCCCCGCGAG 126 12838 GCCAGGCCCCGCCCCCGCGA 125 12839 CGCCAGGCCCCGCCCCCGCG 124 12840 GCGCCAGGCCCCGCCCCCGC 123 12841 GGCGCCAGGCCCCGCCCCCG 122 12842 CGGCGCCAGGCCCCGCCCCC 121 12843 CCGGCGCCAGGCCCCGCCCC 120 12844 GCCGGCGCCAGGCCCCGCCC 119 12845 CGCCGGCGCCAGGCCCCGCC 118 12846 CCGCCGGCGCCAGGCCCCGC 117 12847 ACCGCCGGCGCCAGGCCCCG 116 12848 CACCGCCGGCGCCAGGCCCC 115 12849 CCACCGCCGGCGCCAGGCCC 114 12850 GCCACCGCCGGCGCCAGGCC 113 12851 CGCCACCGCCGGCGCCAGGC 112 12852 ACGCCACCGCCGGCGCCAGG 111 12853 GACGCCACCGCCGGCGCCAG 110 12854 TGACGCCACCGCCGGCGCCA 109 12855 GTGACGCCACCGCCGGCGCC 108 12856 TGTGACGCCACCGCCGGCGC 107 12857 TTGTGACGCCACCGCCGGCG 106 12858 TTTGTGACGCCACCGCCGGC 105 12859 TTTTGTGACGCCACCGCCGG 104 12860 CTTTTGTGACGCCACCGCCG 103 12861 CCTTTTGTGACGCCACCGCC 102 12862 GCCTTTTGTGACGCCACCGC 101 12863 CGCCTTTTGTGACGCCACCG 100 12864 CCGCCTTTTGTGACGCCACC 99 12865 CCCGCCTTTTGTGACGCCAC 98 12866 TCCCGCCTTTTGTGACGCCA 97 12867 GTCCCGCCTTTTGTGACGCC 96 12868 GGTCCCGCCTTTTGTGACGC 95 12869 TGGTCCCGCCTTTTGTGACG 94 12870 GTGGTCCCGCCTTTTGTGAC 93 12871 TGTGGTCCCGCCTTTTGTGA 92 12872 CTGTGGTCCCGCCTTTTGTG 91 12873 ACTGTGGTCCCGCCTTTTGT 90 12874 CACTGTGGTCCCGCCTTTTG 89 12875 CCACTGTGGTCCCGCCTTTT 88 12876 ACCACTGTGGTCCCGCCTTT 87 12877 CACCACTGTGGTCCCGCCTT 86 12878 ACACCACTGTGGTCCCGCCT 85 12879 GACACCACTGTGGTCCCGCC 84 12880 GGACACCACTGTGGTCCCGC 83 12881 CGGACACCACTGTGGTCCCG 82 12882 TCGGACACCACTGTGGTCCC 81 12883 CTCGGACACCACTGTGGTCC 80 12884 TCTCGGACACCACTGTGGTC 79 12885 TTCTCGGACACCACTGTGGT 78 12886 CTTCTCGGACACCACTGTGG 77 12887 ACTTCTCGGACACCACTGTG 76 12888 GACTTCTCGGACACCACTGT 75 12889 TGACTTCTCGGACACCACTG 74 12890 CTGACTTCTCGGACACCACT 73 12891 CCTGACTTCTCGGACACCAC 72 12892 GCCTGACTTCTCGGACACCA 71 12893 TGCCTGACTTCTCGGACACC 70 12894 GTGCCTGACTTCTCGGACAC 69 12895 CGTGCCTGACTTCTCGGACA 68 12896 ACGTGCCTGACTTCTCGGAC 67 12897 TACGTGCCTGACTTCTCGGA 66 12898 CTACGTGCCTGACTTCTCGG 65 12899 GCTACGTGCCTGACTTCTCG 64 12900 AGCTACGTGCCTGACTTCTC 63 12901 GAGCTACGTGCCTGACTTCT 62 12902 TGAGCTACGTGCCTGACTTC 61 12903 CTGAGCTACGTGCCTGACTT 60 12904 GCTGAGCTACGTGCCTGACT 59 12905 CGCTGAGCTACGTGCCTGAC 58 12906 CCGCTGAGCTACGTGCCTGA 57 12907 GCCGCTGAGCTACGTGCCTG 56 12908 CGCCGCTGAGCTACGTGCCT 55 12909 CCGCCGCTGAGCTACGTGCC 54 12910 GCCGCCGCTGAGCTACGTGC 53 12911 GGCCGCCGCTGAGCTACGTG 52 12912 CGCCTGCCTCGGCTCGACCCCCGCAG 177 12913 GCCTGCCTCGGCTCGACCCC 178 12914 CCTGCCTCGGCTCGACCCCC 179 12915 CTGCCTCGGCTCGACCCCCG 180 12916 TGCCTCGGCTCGACCCCCGC 181 12917 GCCTCGGCTCGACCCCCGCA 182 12918 CCTCGGCTCGACCCCCGCAG 183 12919 CTCGGCTCGACCCCCGCAGG 184 12920 TCGGCTCGACCCCCGCAGGG 185 12921 CGGCTCGACCCCCGCAGGGC 186 12922 GGCTCGACCCCCGCAGGGCA 187 12923 GCTCGACCCCCGCAGGGCAG 188 12924 CTCGACCCCCGCAGGGCAGG 189 12925 TCGACCCCCGCAGGGCAGGA 190 12926 CGACCCCCGCAGGGCAGGAC 191 12927 GACCCCCGCAGGGCAGGACC 192 12928 ACCCCCGCAGGGCAGGACCC 193 12929 CCCCCGCAGGGCAGGACCCT 194 12930 CCCCGCAGGGCAGGACCCTG 195 12931 CCCGCAGGGCAGGACCCTGG 196 12932 CCGCAGGGCAGGACCCTGGG 197 12933 CGCAGGGCAGGACCCTGGGC 198 12934 GCAGGGCAGGACCCTGGGCG 199 12935 CAGGGCAGGACCCTGGGCGA 200 12936 AGGGCAGGACCCTGGGCGAC 201 12937 GGGCAGGACCCTGGGCGACT 202 12938 GGCAGGACCCTGGGCGACTC 203 12939 GCAGGACCCTGGGCGACTCC 204 12940 CAGGACCCTGGGCGACTCCG 205 12941 AGGACCCTGGGCGACTCCGC 206 12942 GGACCCTGGGCGACTCCGCC 207 12943 GACCCTGGGCGACTCCGCCC 208 12944 ACCCTGGGCGACTCCGCCCG 209 12945 CCCTGGGCGACTCCGCCCGT 210 12946 CCTGGGCGACTCCGCCCGTT 211 12947 CTGGGCGACTCCGCCCGTTC 212 12948 TGGGCGACTCCGCCCGTTCC 213 12949 GGGCGACTCCGCCCGTTCCT 214 12950 GGCGACTCCGCCCGTTCCTC 215 12951 GCGACTCCGCCCGTTCCTCC 216 12952 CGACTCCGCCCGTTCCTCCA 217 12953 GACTCCGCCCGTTCCTCCAG 218 12954 ACTCCGCCCGTTCCTCCAGC 219 12955 CTCCGCCCGTTCCTCCAGCA 220 12956 TCCGCCCGTTCCTCCAGCAA 221 12957 CCGCCCGTTCCTCCAGCAAC 222 12958 CGCCCGTTCCTCCAGCAACC 223 12959 GCCCGTTCCTCCAGCAACCG 224 12960 CCCGTTCCTCCAGCAACCGC 225 12961 CCGTTCCTCCAGCAACCGCC 226 12962 CGTTCCTCCAGCAACCGCCG 227 12963 GTTCCTCCAGCAACCGCCGC 228 12964 TTCCTCCAGCAACCGCCGCT 229 12965 TCCTCCAGCAACCGCCGCTA 230 12966 CCTCCAGCAACCGCCGCTAA 231 12967 CTCCAGCAACCGCCGCTAAG 232 12968 TCCAGCAACCGCCGCTAAGC 233 12969 CCAGCAACCGCCGCTAAGCC 234 12970 CAGCAACCGCCGCTAAGCCC 235 12971 AGCAACCGCCGCTAAGCCCG 236 12972 GCAACCGCCGCTAAGCCCGG 237 12973 CAACCGCCGCTAAGCCCGGC 238 12974 AACCGCCGCTAAGCCCGGCG 239 12975 ACCGCCGCTAAGCCCGGCGC 240 12976 CCGCCGCTAAGCCCGGCGCA 241 12977 CGCCGCTAAGCCCGGCGCAC 242 12978 GCCGCTAAGCCCGGCGCACC 243 12979 CCGCTAAGCCCGGCGCACCG 244 12980 CGCTAAGCCCGGCGCACCGC 245 12981 GCTAAGCCCGGCGCACCGCT 246 12982 CTAAGCCCGGCGCACCGCTC 247 12983 TAAGCCCGGCGCACCGCTCC 248 12984 AAGCCCGGCGCACCGCTCCA 249 12985 AGCCCGGCGCACCGCTCCAA 250 12986 GCCCGGCGCACCGCTCCAAC 251 12987 CCCGGCGCACCGCTCCAACC 252 12988 CCGGCGCACCGCTCCAACCT 253 12989 CGGCGCACCGCTCCAACCTG 254 12990 GGCGCACCGCTCCAACCTGT 255 12991 GCGCACCGCTCCAACCTGTT 256 12992 CGCACCGCTCCAACCTGTTC 257 12993 GCACCGCTCCAACCTGTTCT 258 12994 CACCGCTCCAACCTGTTCTC 259 12995 ACCGCTCCAACCTGTTCTCC 260 12996 CCGCTCCAACCTGTTCTCCA 261 12997 CGCTCCAACCTGTTCTCCAC 262 12998 CCGCCTGCCTCGGCTCGACC 176 12999 ACCGCCTGCCTCGGCTCGAC 175 13000 CACCGCCTGCCTCGGCTCGA 174 13001 TCACCGCCTGCCTCGGCTCG 173 13002 GTCACCGCCTGCCTCGGCTC 172 13003 AGTCACCGCCTGCCTCGGCT 171 13004 AAGTCACCGCCTGCCTCGGC 170 13005 GAAGTCACCGCCTGCCTCGG 169 13006 GGAAGTCACCGCCTGCCTCG 168 13007 GGGAAGTCACCGCCTGCCTC 167 13008 GGGGAAGTCACCGCCTGCCT 166 13009 TGGGGAAGTCACCGCCTGCC 165 13010 GTGGGGAAGTCACCGCCTGC 164 13011 AGTGGGGAAGTCACCGCCTG 163 13012 GAGTGGGGAAGTCACCGCCT 162 13013 CGAGTGGGGAAGTCACCGCC 161 13014 CCGAGTGGGGAAGTCACCGC 160 13015 CCCGAGTGGGGAAGTCACCG 159 13016 CCCCGAGTGGGGAAGTCACC 158 13017 GCCCCGAGTGGGGAAGTCAC 157 13018 CGCCCCGAGTGGGGAAGTCA 156 13019 CCGCCCCGAGTGGGGAAGTC 155 13020 TCCGCCCCGAGTGGGGAAGT 154 13021 CTCCGCCCCGAGTGGGGAAG 153 13022 GCTCCGCCCCGAGTGGGGAA 152 13023 GGCTCCGCCCCGAGTGGGGA 151 13024 CGGCTCCGCCCCGAGTGGGG 150 13025 GCGGCTCCGCCCCGAGTGGG 149 13026 TGCGGCTCCGCCCCGAGTGG 148 13027 CTGCGGCTCCGCCCCGAGTG 147 13028 GCTGCGGCTCCGCCCCGAGT 146 13029 GGCTGCGGCTCCGCCCCGAG 145 13030 AGGCTGCGGCTCCGCCCCGA 144 13031 GAGGCTGCGGCTCCGCCCCG 143 13032 CGAGGCTGCGGCTCCGCCCC 142 13033 GCGAGGCTGCGGCTCCGCCC 141 13034 CGCGAGGCTGCGGCTCCGCC 140 13035 CCGCGAGGCTGCGGCTCCGC 139 13036 CCCGCGAGGCTGCGGCTCCG 138 13037 CCCCGCGAGGCTGCGGCTCC 137 13038 CCCCCGCGAGGCTGCGGCTC 136 13039 GCCCCCGCGAGGCTGCGGCT 135 13040 CGCCCCCGCGAGGCTGCGGC 134 13041 CCGCCCCCGCGAGGCTGCGG 133 13042 CCCGCCCCCGCGAGGCTGCG 132 13043 CCCCGCCCCCGCGAGGCTGC 131 13044 GCCCCGCCCCCGCGAGGCTG 130 13045 GGCCCCGCCCCCGCGAGGCT 129 13046 AGGCCCCGCCCCCGCGAGGC 128 13047 CAGGCCCCGCCCCCGCGAGG 127 13048 CCAGGCCCCGCCCCCGCGAG 126 13049 GCCAGGCCCCGCCCCCGCGA 125 13050 CGCCAGGCCCCGCCCCCGCG 124 13051 GCGCCAGGCCCCGCCCCCGC 123 13052 GGCGCCAGGCCCCGCCCCCG 122 13053 CGGCGCCAGGCCCCGCCCCC 121 13054 CCGGCGCCAGGCCCCGCCCC 120 13055 GCCGGCGCCAGGCCCCGCCC 119 13056 CGCCGGCGCCAGGCCCCGCC 118 13057 CCGCCGGCGCCAGGCCCCGC 117 13058 ACCGCCGGCGCCAGGCCCCG 116 13059 CACCGCCGGCGCCAGGCCCC 115 13060 CCACCGCCGGCGCCAGGCCC 114 13061 GCCACCGCCGGCGCCAGGCC 113 13062 CGCCACCGCCGGCGCCAGGC 112 13063 ACGCCACCGCCGGCGCCAGG 111 13064 GACGCCACCGCCGGCGCCAG 110 13065 TGACGCCACCGCCGGCGCCA 109 13066 GTGACGCCACCGCCGGCGCC 108 13067 TGTGACGCCACCGCCGGCGC 107 13068 TTGTGACGCCACCGCCGGCG 106 13069 TTTGTGACGCCACCGCCGGC 105 13070 TTTTGTGACGCCACCGCCGG 104 13071 CTTTTGTGACGCCACCGCCG 103 13072 CCTTTTGTGACGCCACCGCC 102 13073 GCCTTTTGTGACGCCACCGC 101 13074 CGCCTTTTGTGACGCCACCG 100 13075 CCGCCTTTTGTGACGCCACC 99 13076 CCCGCCTTTTGTGACGCCAC 98 13077 TCCCGCCTTTTGTGACGCCA 97 13078 GTCCCGCCTTTTGTGACGCC 96 13079 GGTCCCGCCTTTTGTGACGC 95 13080 TGGTCCCGCCTTTTGTGACG 94 13081 GTGGTCCCGCCTTTTGTGAC 93 13082 TGTGGTCCCGCCTTTTGTGA 92 13083 CTGTGGTCCCGCCTTTTGTG 91 13084 ACTGTGGTCCCGCCTTTTGT 90 13085 CACTGTGGTCCCGCCTTTTG 89 13086 CCACTGTGGTCCCGCCTTTT 88 13087 ACCACTGTGGTCCCGCCTTT 87 13088 CACCACTGTGGTCCCGCCTT 86 13089 ACACCACTGTGGTCCCGCCT 85 13090 GACACCACTGTGGTCCCGCC 84 13091 GGACACCACTGTGGTCCCGC 83 13092 CGGACACCACTGTGGTCCCG 82 13093 TCGGACACCACTGTGGTCCC 81 13094 CTCGGACACCACTGTGGTCC 80 13095 TCTCGGACACCACTGTGGTC 79 13096 TTCTCGGACACCACTGTGGT 78 13097 CTTCTCGGACACCACTGTGG 77 13098 ACTTCTCGGACACCACTGTG 76 13099 GACTTCTCGGACACCACTGT 75 13100 TGACTTCTCGGACACCACTG 74 13101 CTGACTTCTCGGACACCACT 73 13102 CCTGACTTCTCGGACACCAC 72 13103 GCCTGACTTCTCGGACACCA 71 13104 TGCCTGACTTCTCGGACACC 70 13105 GTGCCTGACTTCTCGGACAC 69 13106 CGTGCCTGACTTCTCGGACA 68 13107 ACGTGCCTGACTTCTCGGAC 67 13108 TACGTGCCTGACTTCTCGGA 66 13109 CTACGTGCCTGACTTCTCGG 65 13110 GCTACGTGCCTGACTTCTCG 64 13111 AGCTACGTGCCTGACTTCTC 63 13112 GAGCTACGTGCCTGACTTCT 62 13113 TGAGCTACGTGCCTGACTTC 61 13114 CTGAGCTACGTGCCTGACTT 60 13115 GCTGAGCTACGTGCCTGACT 59 13116 CGCTGAGCTACGTGCCTGAC 58 13117 CCGCTGAGCTACGTGCCTGA 57 13118 GCCGCTGAGCTACGTGCCTG 56 13119 CGCCGCTGAGCTACGTGCCT 55 13120 CCGCCGCTGAGCTACGTGCC 54 13121 GCCGCCGCTGAGCTACGTGC 53 13122 GGCCGCCGCTGAGCTACGTG 52 13123 CGGCTAGAAATCGGCCTGTTCCGGCCTCGCCT 286 13124 GGCTAGAAATCGGCCTGTTC 287 13125 GCTAGAAATCGGCCTGTTCC 288 13126 CTAGAAATCGGCCTGTTCCG 289 13127 TAGAAATCGGCCTGTTCCGG 290 13128 AGAAATCGGCCTGTTCCGGC 291 13129 GAAATCGGCCTGTTCCGGCC 292 13130 AAATCGGCCTGTTCCGGCCT 293 13131 AATCGGCCTGTTCCGGCCTC 294 13132 ATCGGCCTGTTCCGGCCTCG 295 13133 TCGGCCTGTTCCGGCCTCGC 296 13134 CGGCCTGTTCCGGCCTCGCC 297 13135 GGCCTGTTCCGGCCTCGCCT 298 13136 GCCTGTTCCGGCCTCGCCTC 299 13137 CCTGTTCCGGCCTCGCCTCG 300 13138 CTGTTCCGGCCTCGCCTCGG 301 13139 TGTTCCGGCCTCGCCTCGGG 302 13140 GTTCCGGCCTCGCCTCGGGT 303 13141 TTCCGGCCTCGCCTCGGGTC 304 13142 TCCGGCCTCGCCTCGGGTCT 305 13143 CCGGCCTCGCCTCGGGTCTT 306 13144 CGGCCTCGCCTCGGGTCTTT 307 13145 GGCCTCGCCTCGGGTCTTTC 308 13146 GCCTCGCCTCGGGTCTTTCT 309 13147 CCTCGCCTCGGGTCTTTCTT 310 13148 CTCGCCTCGGGTCTTTCTTA 311 13149 TCGCCTCGGGTCTTTCTTAG 312 13150 CGCCTCGGGTCTTTCTTAGT 313 13151 GCCTCGGGTCTTTCTTAGTC 314 13152 CCTCGGGTCTTTCTTAGTCC 315 13153 CTCGGGTCTTTCTTAGTCCT 316 13154 TCGGGTCTTTCTTAGTCCTT 317 13155 CGGGTCTTTCTTAGTCCTTT 318 13156 GCGGCTAGAAATCGGCCTGT 285 13157 GGCGGCTAGAAATCGGCCTG 284 13158 TGGCGGCTAGAAATCGGCCT 283 13159 TTGGCGGCTAGAAATCGGCC 282 13160 CTTGGCGGCTAGAAATCGGC 281 13161 ACTTGGCGGCTAGAAATCGG 280 13162 CACTTGGCGGCTAGAAATCG 279 13163 CCACTTGGCGGCTAGAAATC 278 13164 TCCACTTGGCGGCTAGAAAT 277 13165 CTCCACTTGGCGGCTAGAAA 276 13166 TCTCCACTTGGCGGCTAGAA 275 13167 TTCTCCACTTGGCGGCTAGA 274 13168 GTTCTCCACTTGGCGGCTAG 273 13169 TGTTCTCCACTTGGCGGCTA 272 13170 CTGTTCTCCACTTGGCGGCT 271 13171 CCTGTTCTCCACTTGGCGGC 270 13172 ACCTGTTCTCCACTTGGCGG 269 13173 AACCTGTTCTCCACTTGGCG 268 13174 CGGGGGTGGGGATGCGGCGGTGAACCCG 377 13175 CGCGGCAGGTGAGAGGGGAGCTGCCCCTGCG 558 13176 CGCGTGCACGTGTGTCCACATGAGTGC 3650 13177 GCGTGCACGTGTGTCCACAT 3651 13178 CGTGCACGTGTGTCCACATG 3652 13179 GTGCACGTGTGTCCACATGA 3653 13180 TGCACGTGTGTCCACATGAG 3654 13181 GCACGTGTGTCCACATGAGT 3655 13182 CACGTGTGTCCACATGAGTG 3656 13183 ACGTGTGTCCACATGAGTGC 3657 13184 CGTGTGTCCACATGAGTGCT 3658 13185 GCGCGTGCACGTGTGTCCAC 3649 13186 TGCGCGTGCACGTGTGTCCA 3648 13187 GTGCGCGTGCACGTGTGTCC 3647 13188 TGTGCGCGTGCACGTGTGTC 3646 13189 GTGTGCGCGTGCACGTGTGT 3645 13190 TGTGTGCGCGTGCACGTGTG 3644 13191 GTGTGTGCGCGTGCACGTGT 3643 13192 TGTGTGTGCGCGTGCACGTG 3642 13193 ATGTGTGTGCGCGTGCACGT 3641 13194 CATGTGTGTGCGCGTGCACG 3640 13195 CCATGTGTGTGCGCGTGCAC 3639 13196 TCCATGTGTGTGCGCGTGCA 3638 13197 GTCCATGTGTGTGCGCGTGC 3637 13198 TGTCCATGTGTGTGCGCGTG 3636 13199 GTGTCCATGTGTGTGCGCGT 3635 13200 TGTGTCCATGTGTGTGCGCG 3634 13201 GTGTGTCCATGTGTGTGCGC 3633 13202 TGTGTGTCCATGTGTGTGCG 3632 13203 CGCCACCGCCGGCGCCAGGCCCCGCC 112 13204 GCCACCGCCGGCGCCAGGCC 113 13205 CCACCGCCGGCGCCAGGCCC 114 13206 CACCGCCGGCGCCAGGCCCC 115 13207 ACCGCCGGCGCCAGGCCCCG 116 13208 CCGCCGGCGCCAGGCCCCGC 117 13209 CGCCGGCGCCAGGCCCCGCC 118 13210 GCCGGCGCCAGGCCCCGCCC 119 13211 CCGGCGCCAGGCCCCGCCCC 120 13212 CGGCGCCAGGCCCCGCCCCC 121 13213 GGCGCCAGGCCCCGCCCCCG 122 13214 GCGCCAGGCCCCGCCCCCGC 123 13215 CGCCAGGCCCCGCCCCCGCG 124 13216 GCCAGGCCCCGCCCCCGCGA 125 13217 CCAGGCCCCGCCCCCGCGAG 126 13218 CAGGCCCCGCCCCCGCGAGG 127 13219 AGGCCCCGCCCCCGCGAGGC 128 13220 GGCCCCGCCCCCGCGAGGCT 129 13221 GCCCCGCCCCCGCGAGGCTG 130 13222 CCCCGCCCCCGCGAGGCTGC 131 13223 CCCGCCCCCGCGAGGCTGCG 132 13224 CCGCCCCCGCGAGGCTGCGG 133 13225 CGCCCCCGCGAGGCTGCGGC 134 13226 GCCCCCGCGAGGCTGCGGCT 135 13227 CCCCCGCGAGGCTGCGGCTC 136 13228 CCCCGCGAGGCTGCGGCTCC 137 13229 CCCGCGAGGCTGCGGCTCCG 138 13230 CCGCGAGGCTGCGGCTCCGC 139 13231 CGCGAGGCTGCGGCTCCGCC 140 13232 GCGAGGCTGCGGCTCCGCCC 141 13233 CGAGGCTGCGGCTCCGCCCC 142 13234 GAGGCTGCGGCTCCGCCCCG 143 13235 AGGCTGCGGCTCCGCCCCGA 144 13236 GGCTGCGGCTCCGCCCCGAG 145 13237 GCTGCGGCTCCGCCCCGAGT 146 13238 CTGCGGCTCCGCCCCGAGTG 147 13239 TGCGGCTCCGCCCCGAGTGG 148 13240 GCGGCTCCGCCCCGAGTGGG 149 13241 CGGCTCCGCCCCGAGTGGGG 150 13242 GGCTCCGCCCCGAGTGGGGA 151 13243 GCTCCGCCCCGAGTGGGGAA 152 13244 CTCCGCCCCGAGTGGGGAAG 153 13245 TCCGCCCCGAGTGGGGAAGT 154 13246 CCGCCCCGAGTGGGGAAGTC 155 13247 CGCCCCGAGTGGGGAAGTCA 156 13248 GCCCCGAGTGGGGAAGTCAC 157 13249 CCCCGAGTGGGGAAGTCACC 158 13250 CCCGAGTGGGGAAGTCACCG 159 13251 CCGAGTGGGGAAGTCACCGC 160 13252 CGAGTGGGGAAGTCACCGCC 161 13253 GAGTGGGGAAGTCACCGCCT 162 13254 AGTGGGGAAGTCACCGCCTG 163 13255 GTGGGGAAGTCACCGCCTGC 164 13256 TGGGGAAGTCACCGCCTGCC 165 13257 GGGGAAGTCACCGCCTGCCT 166 13258 GGGAAGTCACCGCCTGCCTC 167 13259 GGAAGTCACCGCCTGCCTCG 168 13260 GAAGTCACCGCCTGCCTCGG 169 13261 AAGTCACCGCCTGCCTCGGC 170 13262 AGTCACCGCCTGCCTCGGCT 171 13263 GTCACCGCCTGCCTCGGCTC 172 13264 TCACCGCCTGCCTCGGCTCG 173 13265 CACCGCCTGCCTCGGCTCGA 174 13266 ACCGCCTGCCTCGGCTCGAC 175 13267 CCGCCTGCCTCGGCTCGACC 176 13268 CGCCTGCCTCGGCTCGACCC 177 13269 GCCTGCCTCGGCTCGACCCC 178 13270 CCTGCCTCGGCTCGACCCCC 179 13271 CTGCCTCGGCTCGACCCCCG 180 13272 TGCCTCGGCTCGACCCCCGC 181 13273 GCCTCGGCTCGACCCCCGCA 182 13274 CCTCGGCTCGACCCCCGCAG 183 13275 CTCGGCTCGACCCCCGCAGG 184 13276 TCGGCTCGACCCCCGCAGGG 185 13277 CGGCTCGACCCCCGCAGGGC 186 13278 GGCTCGACCCCCGCAGGGCA 187 13279 GCTCGACCCCCGCAGGGCAG 188 13280 CTCGACCCCCGCAGGGCAGG 189 13281 TCGACCCCCGCAGGGCAGGA 190 13282 CGACCCCCGCAGGGCAGGAC 191 13283 GACCCCCGCAGGGCAGGACC 192 13284 ACCCCCGCAGGGCAGGACCC 193 13285 CCCCCGCAGGGCAGGACCCT 194 13286 CCCCGCAGGGCAGGACCCTG 195 13287 CCCGCAGGGCAGGACCCTGG 196 13288 CCGCAGGGCAGGACCCTGGG 197 13289 CGCAGGGCAGGACCCTGGGC 198 13290 GCAGGGCAGGACCCTGGGCG 199 13291 CAGGGCAGGACCCTGGGCGA 200 13292 AGGGCAGGACCCTGGGCGAC 201 13293 GGGCAGGACCCTGGGCGACT 202 13294 GGCAGGACCCTGGGCGACTC 203 13295 GCAGGACCCTGGGCGACTCC 204 13296 CAGGACCCTGGGCGACTCCG 205 13297 AGGACCCTGGGCGACTCCGC 206 13298 GGACCCTGGGCGACTCCGCC 207 13299 GACCCTGGGCGACTCCGCCC 208 13300 ACCCTGGGCGACTCCGCCCG 209 13301 CCCTGGGCGACTCCGCCCGT 210 13302 CCTGGGCGACTCCGCCCGTT 211 13303 CTGGGCGACTCCGCCCGTTC 212 13304 TGGGCGACTCCGCCCGTTCC 213 13305 GGGCGACTCCGCCCGTTCCT 214 13306 GGCGACTCCGCCCGTTCCTC 215 13307 GCGACTCCGCCCGTTCCTCC 216 13308 CGACTCCGCCCGTTCCTCCA 217 13309 GACTCCGCCCGTTCCTCCAG 218 13310 ACTCCGCCCGTTCCTCCAGC 219 13311 CTCCGCCCGTTCCTCCAGCA 220 13312 TCCGCCCGTTCCTCCAGCAA 221 13313 CCGCCCGTTCCTCCAGCAAC 222 13314 CGCCCGTTCCTCCAGCAACC 223 13315 GCCCGTTCCTCCAGCAACCG 224 13316 CCCGTTCCTCCAGCAACCGC 225 13317 CCGTTCCTCCAGCAACCGCC 226 13318 CGTTCCTCCAGCAACCGCCG 227 13319 GTTCCTCCAGCAACCGCCGC 228 13320 TTCCTCCAGCAACCGCCGCT 229 13321 TCCTCCAGCAACCGCCGCTA 230 13322 CCTCCAGCAACCGCCGCTAA 231 13323 CTCCAGCAACCGCCGCTAAG 232 13324 TCCAGCAACCGCCGCTAAGC 233 13325 CCAGCAACCGCCGCTAAGCC 234 13326 CAGCAACCGCCGCTAAGCCC 235 13327 AGCAACCGCCGCTAAGCCCG 236 13328 GCAACCGCCGCTAAGCCCGG 237 13329 CAACCGCCGCTAAGCCCGGC 238 13330 AACCGCCGCTAAGCCCGGCG 239 13331 ACCGCCGCTAAGCCCGGCGC 240 13332 CCGCCGCTAAGCCCGGCGCA 241 13333 CGCCGCTAAGCCCGGCGCAC 242 13334 GCCGCTAAGCCCGGCGCACC 243 13335 CCGCTAAGCCCGGCGCACCG 244 13336 CGCTAAGCCCGGCGCACCGC 245 13337 GCTAAGCCCGGCGCACCGCT 246 13338 CTAAGCCCGGCGCACCGCTC 247 13339 TAAGCCCGGCGCACCGCTCC 248 13340 AAGCCCGGCGCACCGCTCCA 249 13341 AGCCCGGCGCACCGCTCCAA 250 13342 GCCCGGCGCACCGCTCCAAC 251 13343 CCCGGCGCACCGCTCCAACC 252 13344 CCGGCGCACCGCTCCAACCT 253 13345 CGGCGCACCGCTCCAACCTG 254 13346 GGCGCACCGCTCCAACCTGT 255 13347 GCGCACCGCTCCAACCTGTT 256 13348 CGCACCGCTCCAACCTGTTC 257 13349 GCACCGCTCCAACCTGTTCT 258 13350 CACCGCTCCAACCTGTTCTC 259 13351 ACCGCTCCAACCTGTTCTCC 260 13352 CCGCTCCAACCTGTTCTCCA 261 13353 CGCTCCAACCTGTTCTCCAC 262 13354 ACGCCACCGCCGGCGCCAGG 111 13355 GACGCCACCGCCGGCGCCAG 110 13356 TGACGCCACCGCCGGCGCCA 109 13357 GTGACGCCACCGCCGGCGCC 108 13358 TGTGACGCCACCGCCGGCGC 107 13359 TTGTGACGCCACCGCCGGCG 106 13360 TTTGTGACGCCACCGCCGGC 105 13361 TTTTGTGACGCCACCGCCGG 104 13362 CTTTTGTGACGCCACCGCCG 103 13363 CCTTTTGTGACGCCACCGCC 102 13364 GCCTTTTGTGACGCCACCGC 101 13365 CGCCTTTTGTGACGCCACCG 100 13366 CCGCCTTTTGTGACGCCACC 99 13367 CCCGCCTTTTGTGACGCCAC 98 13368 TCCCGCCTTTTGTGACGCCA 97 13369 GTCCCGCCTTTTGTGACGCC 96 13370 GGTCCCGCCTTTTGTGACGC 95 13371 TGGTCCCGCCTTTTGTGACG 94 13372 GTGGTCCCGCCTTTTGTGAC 93 13373 TGTGGTCCCGCCTTTTGTGA 92 13374 CTGTGGTCCCGCCTTTTGTG 91 13375 ACTGTGGTCCCGCCTTTTGT 90 13376 CACTGTGGTCCCGCCTTTTG 89 13377 CCACTGTGGTCCCGCCTTTT 88 13378 ACCACTGTGGTCCCGCCTTT 87 13379 CACCACTGTGGTCCCGCCTT 86 13380 ACACCACTGTGGTCCCGCCT 85 13381 GACACCACTGTGGTCCCGCC 84 13382 GGACACCACTGTGGTCCCGC 83 13383 CGGACACCACTGTGGTCCCG 82 13384 TCGGACACCACTGTGGTCCC 81 13385 CTCGGACACCACTGTGGTCC 80 13386 TCTCGGACACCACTGTGGTC 79 13387 TTCTCGGACACCACTGTGGT 78 13388 CTTCTCGGACACCACTGTGG 77 13389 ACTTCTCGGACACCACTGTG 76 13390 GACTTCTCGGACACCACTGT 75 13391 TGACTTCTCGGACACCACTG 74 13392 CTGACTTCTCGGACACCACT 73 13393 CCTGACTTCTCGGACACCAC 72 13394 GCCTGACTTCTCGGACACCA 71 13395 TGCCTGACTTCTCGGACACC 70 13396 GTGCCTGACTTCTCGGACAC 69 13397 CGTGCCTGACTTCTCGGACA 68 13398 ACGTGCCTGACTTCTCGGAC 67 13399 TACGTGCCTGACTTCTCGGA 66 13400 CTACGTGCCTGACTTCTCGG 65 13401 GCTACGTGCCTGACTTCTCG 64 13402 AGCTACGTGCCTGACTTCTC 63 13403 GAGCTACGTGCCTGACTTCT 62 13404 TGAGCTACGTGCCTGACTTC 61 13405 CTGAGCTACGTGCCTGACTT 60 13406 GCTGAGCTACGTGCCTGACT 59 13407 CGCTGAGCTACGTGCCTGAC 58 13408 CCGCTGAGCTACGTGCCTGA 57 13409 GCCGCTGAGCTACGTGCCTG 56 13410 CGCCGCTGAGCTACGTGCCT 55 13411 CCGCCGCTGAGCTACGTGCC 54 13412 GCCGCCGCTGAGCTACGTGC 53 13413 GGCCGCCGCTGAGCTACGTG 52 13414 CGCGGCAGGTGAGAGGGGAGCTGCCC 558

Hot Zones (Relative upstream location to gene start site)   1-1880 2150-2240 2420-3050 3230-4130 4310-4400

Examples

In FIG. 64, In MCF7 (human mammary breast cell line), MIF1_(—)1 (329) and MIF1_(—)2 (330) produced statistically significant (P<0.05) inhibition at 10 μM compared to the untreated and negative control values. The MIF1 sequences MIF1_(—)1 (329) and MIF1_(—)2 (330) fit the independent and dependent DNAi motif claims.

The secondary structure for MIF1_(—)1 (329) and MIF1_(—)2 (330) are shown in FIG. 65 and FIG. 66.

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13677) CCATTCTGAGTATCTTCCAAGTGTTAGCTCCTTTAATCCTGGAAAGGACC CCATGAAATTAGTACTTTTATTACCCCTGTTGTACATATGAGAGACTGAG TAAAAGCCGGTGGCTTGTCCAGGGTCACACAGCTAACTGGAATGGCCAGG AGTAGACCTGGTGACCATGGACCCCAGACCTTGATCACTGCACACGCTGC GTCTGGGACCTCGCCTGGTACCTGAGGTCCGTGGCGCGCTGGTGCTGATC ATTCAGAGTGCTCATGGGAAGTGTAGTCTAGAGTCTGTGTGCTTCCTGAT CTCCTTGATCTCCATTTTATTGAGGAGGCCTTTAGGCCACCCGAGGGGTC CAGAGTGACCCTGTGGATTAGCAGTGGAGCTCAGCTTGAGCCAGCGCTCT TCAGGGGTCGTGTTCTGCCCCCATTCTCTGGTTCATTCTGCAGGTAGCAG GGAATCATTGAAGATTAGAGAGAATCAAACACCTGGAGAGAGATGACTCT GCCCGGGGAGCCCAGGCTCCTGTCTGGGTGCACACTCCAGGGCTAGATGG TGACTTCTCAGCTACTCTAGCTTCATAGGCTCATAGTGCATGTGAGCACT CATGTGGACACACGTGCACGCGCACACACATGGACACACACACACACACA CACACCGCTGTCTTTGGAATCAGACCATGAAAATGCTTCCTCAGAGGCCT AGGGGTGAGGAAGCTGAGGTGAGTTGTGCCTCCAGCTGGATGTGCTGGGA TGGGGTGGGAGATGAGGTGGCCACACCTGGGTGGCAGGAACTCTGGGGCA GTGAACCTTCTAACGAACAGATCTGGGATGCTGCCATGAGGAGGAAGAGG GAGTCAGCAGCCATGCCTGCCAATGCCTCCTAGCGCATTTGTCCATGGTT AGCGGATAATTATTGTGTCCCTATGGGTCCCAAGGTGTATTATTTTTTTT TTGCTCTTATAATAAATCAACACAAATTTTTAGCAGCTTCAAACAACACG CATTTATTATCTCACAGTTTCTGTGGGTCAGTAGTCCGGCGTGACATGAC TAGGTCTTCTGTGTAAGGACTCGCATGGCCAAAGTCAAGGTATCTGAAGG GACAAGGGAAAAATCCACTTCCAAGTTCAATCTGGTTGTGAGCAGAATTC AGTTCCTTGTGGTTGTACCATGAGGTCTCTGGTCCCCTTCATCTTCAAAG CCGGTAATGGACATCGAGTGTTTCTCTTGCTTGGAATCTGGCACTCTAGC TGGAGAAAATTATCTGCTTTTAAGAGTTCATGTGATTAGATTGGGTGTAC CCAGATGCTCCATGCTAATCTCCCTATTATGCACAGATGCATAATCCTAA TTGCATCTGTGAAGTGCTTTTTGCCAGGTAACATGGCATACTTGTAGGTT CCAGGGATTAGTGCTTGTCCTCCCCCTGCTATTCTTTAGTGGGCAGGGGG TCATCTGCCTACCACGGAGGTAAGGGGTCAGGAGGTATGCATACAGCAAT GCCCAAAAAGAGACTGTCCCCACTGGGATGGAGTTTACCGCCTAGACATG CAGTCTTAACTCAGAAATATGGAGATAGCCTCGAAGGACAGGACAGGTAC TGGGCACGTGTGGGAATGGACCAAGCCAGGTGCTCCGGGGGCTTTCCCAA GGAACTAAGGCTGAGCCAAGAACTGAAGGATGAGTTGGAGTCAGATGAGG GAAAATGTGGGCAAACTGGATTTCAGAACCAACCCCCAACCCTGGAGCCA GGAGCCATGGTACTGAAGGACAGTGCGCCATAACTCAGAGAACCAGGGAG GGTTGGCGGAGGCTCACAGGGACCGGGTTACCCCAGGGCCTTGTGACAGT ACTACCCCTAGTATCAGAGGAGACTGTCATTGGCATTTAGGCCACTTGGT GCTCATAACACCTCTATGTCAGGTGAACACTATTGTCATCCCCAAATTAC AGATGGGGAAAGTGAGCCAAATGTCCATGCTAGTAAGAGGCAAATCATAT CACTTCTTTGGGTACCCTTCTAGAAGGATGAGGCTGACTGCCACTGGAAA CAGCTGGGGAGGGTACAAGGAGATGACAAGTGGCTCAGAGGCTGTCCTGG CTATAAGAATTAAAGAGGAAAGAAACACCAAGGGTGGCTCGACAGTCAAC AAGGACAGGTTTATTTTGGAAAACAAACTTGAGAGGGGCTTCTGGCCAAG TTAGGTCAGAGCCACACTCTCTTACAAACTAAGGATATTTAAGGGTTTTG GAGGGGGTTCTTATCATAGGTTCTGAATGTTTCTGTGTGAGGGAAAGTTT ATTGCGGGGATGGAATGTCTCTGGTCAGAAGGGAGGCTGTCTCCGGGTTG GCATGTTTCTGGTCAGAGAAGGGTTTATCTTAGGGTTGGAATGTTTCTGG TTATGCTGACATTAGCTATTAGGCTGATATTTTCGGGCTGGATTTAGGCG GCTTTTAATTAAGGGGGAACTTAGAATGGTGGTGTTTGTTCAAGATGGCA ATGCTCCTGCTCCGTCACTGGCCAGGTAAGGCAACCCTTTGTTATGGTAA CAACCTGAGATTGGCAGGGGCTCACCTCCAGGGGCAGCTCATGTGCTTGC TGGCGAGGCTGCACCTTGTCATTCAGGTTCACAGGGCACAGGTCAACCAG GCCCTGGCTCTTCAGTCTTCTGCCTGGAGTGACTTATGTAATTCTGCTCA GCTTTCATAGGGCACAGGGAGTCGGGGCTAACTCTGCTGCCTGGGGCTGG AAACAGACTCCTCCCTTGAGGAGCAGCAGTCCACCATAGGGAAGTCACAG TGGTCCAGGCCAAAGGGGATGCAGGTAGTGTAGACTAGGCGGTAGTTCAG GGAATGGAGAGAAGTGGGAATAAAGGGATAGTGAAAGGAAGCATATTTTA CTGGCAGGTGATGAGGTGTAGGAGGACAAGTCATACATTTGGACTTTACA GAGCAGTGGACACTCAGTCAGCTGCTGTCAGCGCCTGGGACTTAGGGGAG TGCCCCTGGCTGGAGACATGGTATGGAGTGCCATCAGTTAGGGAGCCCTG GGCACAGGTAAGAGAAGGTGTGACACCAGGAGGGAAAGAGTCTGGGGCCC AGCTGCAGGAACCAATACCCATAGGCTATTTGTATAAATGGGCCATGGGG CCTCCCAGCTGGAGGCTGGCTGGTGCCACGAGGGTCCCACAGGCATGGGT GTCCTTCCTATATCACATGGCCTTCACTGAGACTGGTATATGGATTGCAC CTATCAGAGACCAAGGACAGGACCTCCCTGGAAATCTCTGAGGACCTGGC CTGTGATCCAGTTGCTGCCTTGTCCTCTTCCTGCTATGTCATGGCTTATC TTCTTTCACCCATTCATTCATTCATTCATTCAGCAGTATTAGTCAATGTC TCTTGTATGCCTGGCACCTGCTAGATGGTCCCCGAGTTTACCATTAGTGG AAAAGACATTTAAGAAATTCACCAAGGGCTCTATGAGAGGCCATACACGG TGGACCTGACTAGGGTGTGGCTTCCCTGAGGAGCTGAAGTTGCCCAGAGG CCCAGAGAAGGGGAGCTGAGCACGTTTGAACCACTGAACCTGCTCTGGAC CTCGCCTCCTTCCCTTCGGTGCCTCCCAGCATCCTATCCTCTTTAAAGAG CAGGGGTTCAGGGAAGTTCCCTGGATGGTGATTCGCAGGGGCAGCTCCCC TCTCACCTGCCGCGATGACTACCCCGCCCCATCTCAAACACACAAGCTCA CGCATGCGGGACTGGAGCCCTTGAGGACATGTGGCCCAAAGACAGGAGGT ACAGGGGCTCAGTGCGTGCAGTGGAATGAACTGGGCTTCATCTCTGGAAG GGTAAGGGGCCATCTTCCGGGTTCACCGCCGCATCCCCACCCCCGGCACA GCGCCTCCTGGCGACTAACATCGGTGACTTAGTGAAAGGACTAAGAAAGA CCCGAGGCGAGGCCGGAACAGGCCGATTTCTAGCCGCCAAGTGGAGAACA GGTTGGAGCGGTGCGCCGGGCTTAGCGGCGGTTGCTGGAGGAACGGGCGG AGTCGCCCAGGGTCCTGCCCTGCGGGGGTCGAGCCGAGGCAGGCGGTGAC TTCCCCACTCGGGGCGGAGCCGCAGCCTCGCGGGGGCGGGGCCTGGCGCC GGCGGTGGCGTCACAAAAGGCGGGACCACAGTGGTGTCCGAGAAGTCAGG CACGTAGCTCAGCGGCGGCCGCGGCGCGTGCGTCTGTGCCTCTGCGCGGG TCTCCTGGTCCTTCTGCCATC ATG

ERBB2

ERBB2 (also known as HER2/meu and CD340) is a receptor tyrosine kinase protein and member of the epidermal growth factor receptor family. ERBB2 contains extracellular, transmembrane, and intracellular domains. Ligand binding causes dimerization which activates downstream signaling pathways leading to proliferation, cell cycle progression, and cell survival promotion. ERBB2 is commonly associated with breast cancer where the gene is amplified or the protein is overexpressed leading to dysregulation of cell proliferation and survival. ERBB2 has also been associated with other cancers including lung and colorectal cancer.

Protein: ERBB2 (HER2) Gene: ERBB2 (Homo sapiens, chromosome 17, 37844167-37884915 [NCBI Reference Sequence: NC_(—)000017.10]; start site location: 37855813; strand: positive)

Gene Identification GeneID 2064 HGNC 3430 MIM 164870

Targeted Sequences Relative upstream location to gene Sequence Design start ID No: ID Sequence (5′-3′) site 13415 CGGGAAGAGGATGCGCTGACCTGGC 2571 13416 CACGCCCTGGGGAGGAGGCTCGAGAGG 3267 13437 CGAGAGGGGCCGAGCCTCTGAAAAA 3287 13452 CGTCTGGTCCACAGTCCGATGTCCA 3944

Target Shift Sequences Relative upstream location to Sequence gene ID No: Sequence (5′-3′) start site 13415 CGGGAAGAGGATGCGCTGACCTGGC 2571 13416 CACGCCCTGGGGAGGAGGCTCGAGAGG 3267 13417 ACGCCCTGGGGAGGAGGCTC 3268 13418 CGCCCTGGGGAGGAGGCTCG 3269 13419 GCCCTGGGGAGGAGGCTCGA 3270 13420 CCCTGGGGAGGAGGCTCGAG 3271 13421 TCACGCCCTGGGGAGGAGGC 3266 13422 CTCACGCCCTGGGGAGGAGG 3265 13423 ACTCACGCCCTGGGGAGGAG 3264 13424 AACTCACGCCCTGGGGAGGA 3263 13425 GAACTCACGCCCTGGGGAGG 3262 13426 AGAACTCACGCCCTGGGGAG 3261 13427 CAGAACTCACGCCCTGGGGA 3260 13428 TCAGAACTCACGCCCTGGGG 3259 13429 GTCAGAACTCACGCCCTGGG 3258 13430 GGTCAGAACTCACGCCCTGG 3257 13431 GGGTCAGAACTCACGCCCTG 3256 13432 GGGGTCAGAACTCACGCCCT 3255 13433 TGGGGTCAGAACTCACGCCC 3254 13434 CTGGGGTCAGAACTCACGCC 3253 13435 GCTGGGGTCAGAACTCACGC 3252 13436 AGCTGGGGTCAGAACTCACG 3251 13437 CGAGAGGGGCCGAGCCTCTGAAAAA 3287 13438 GAGAGGGGCCGAGCCTCTGA 3288 13439 AGAGGGGCCGAGCCTCTGAA 3289 13440 GAGGGGCCGAGCCTCTGAAA 3290 13441 AGGGGCCGAGCCTCTGAAAA 3291 13442 GGGGCCGAGCCTCTGAAAAA 3292 13443 GGGCCGAGCCTCTGAAAAAG 3293 13444 GGCCGAGCCTCTGAAAAAGA 3294 13445 GCCGAGCCTCTGAAAAAGAA 3295 13446 CCGAGCCTCTGAAAAAGAAT 3296 13447 CGAGCCTCTGAAAAAGAATG 3297 13448 TCGAGAGGGGCCGAGCCTCT 3286 13449 CTCGAGAGGGGCCGAGCCTC 3285 13450 GCTCGAGAGGGGCCGAGCCT 3284 13451 GGCTCGAGAGGGGCCGAGCC 3283 13452 CGTCTGGTCCACAGTCCGATGTCCA 3944 13453 GTCTGGTCCACAGTCCGATG 3945 13454 TCTGGTCCACAGTCCGATGT 3946 13455 CTGGTCCACAGTCCGATGTC 3947 13456 TGGTCCACAGTCCGATGTCC 3948 13457 GGTCCACAGTCCGATGTCCA 3949 13458 GTCCACAGTCCGATGTCCAG 3950 13459 TCCACAGTCCGATGTCCAGG 3951 13460 CCACAGTCCGATGTCCAGGC 3952 13461 CACAGTCCGATGTCCAGGCC 3953 13462 ACAGTCCGATGTCCAGGCCA 3954 13463 CAGTCCGATGTCCAGGCCAC 3955 13464 AGTCCGATGTCCAGGCCACA 3956 13465 GTCCGATGTCCAGGCCACAA 3957 13466 TCCGATGTCCAGGCCACAAA 3958 13467 CCGATGTCCAGGCCACAAAC 3959 13468 CGATGTCCAGGCCACAAACT 3960 13469 TCGTCTGGTCCACAGTCCGA 3943 13470 GTCGTCTGGTCCACAGTCCG 3942 13471 AGTCGTCTGGTCCACAGTCC 3941 13472 GAGTCGTCTGGTCCACAGTC 3940 13473 GGAGTCGTCTGGTCCACAGT 3939 13474 AGGAGTCGTCTGGTCCACAG 3938 13475 GAGGAGTCGTCTGGTCCACA 3937 13476 GGAGGAGTCGTCTGGTCCAC 3936 13477 GGGAGGAGTCGTCTGGTCCA 3935 13478 CGGGAGGAGTCGTCTGGTCC 3934 13479 TCGGGAGGAGTCGTCTGGTC 3933 13480 ATCGGGAGGAGTCGTCTGGT 3932 13481 AATCGGGAGGAGTCGTCTGG 3931 13482 AAATCGGGAGGAGTCGTCTG 3930 13483 GAAATCGGGAGGAGTCGTCT 3929

Hot Zones (Relative upstream location to gene start site) 100-4510

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13678) GGGGGCACCAGTAGAATGGCCAGGACAAACGCAGTGCAGCACAGAGACTC AGACCCTGGCAGCCATGCCTGCGCAGGCAGTGATGAGAGTGACATGTACT GTTGTGGACATGCACAAAAGTGAGGTGAGTCGCAGGACAGAAGAGTGCTT TTTGTTTCAGCAGAGCAGCCTGGGGAGAGATAAAAGCTACTCCTGGGGCC TGGGCCTGCATTCCTGAGATGTGGGTAAGAGGGGCCCAGGGTCAGAGTGT CTGGCAAGCTTGGCTCTGCCCCTTTGCTGTCCTGGAGACTAGGGCTAATC CTGGGCTCAGGGAGTGGCCTCCCCATGGTTAGGATACAAGTGCTCATCAA GGGCCACCCCTAGGAAGGACCAATTTTCCTATCAGAAGCTTCTAAGTTAT CCTCCTTTGGCCCAAAGGGACACCTCAAGCCTACTCTGAGGAACTCTTTC CAATGAACTAATTCCTACAGTCACTTCCCCAGCAACCTGTGCCTCAGCCT CAAGGCACTGTGGGGTAGGCCTCAGTTTGTGGCCTGGACATCGGACTGTG GACCAGACGACTCCTCCCGATTTCTGTTTGTTTTCAGTCCTCTGACCCCA AGCTGGCTGGTGAAGTAGGTAGAGGGAGGAGACTTTGGTGCATGCATACA CACACACACACACACACACACACACACACACACACACACACACACACACA CGTCTCCTGTGCCCCCCAGTCTCCATGGCTGGTCAATGATTGACTGGCAT TTCACAGGCCGCTGGTTGCAGCCCCAGCCTGTTGACTTAGAGGTCACCCT CGGAAGCTAGAGCCCTGTCCTGCCTCTTCAGTGTCAGTGGTCACTCCACT GCCCACAGGCTGGGGTCTTGGGCAAAACACACGCATCTGCCCTGATCTGA GTTTGCTGCCCTCTGTCCCGCAGTCAGCCCCACTCTGTTCCCACTCCCTC TCCCCAGCCCCCTAGCTAGACCCCTCTCACCAGCACCCCTTTCCCTTCCC TGAGGGTCCCCCTCGCTGTCTTTGTCCCTCAGACATCCTCTTTCCTGGGC TCTCCTGCCAGGCCCTGCTGGAGGGACAGTTAAGGAGGAAATCGAATCAG CAGCGCCCACCCCTGCCCCCCTTCCTCTCCTCTTGTCAGACACCAGACGA GGTTTTTTCCTCTGGCTTCCCAGCTCTGAATGGGCTCATTCTTTTTCAGA GGCTCGGCCCCTCTCGAGCCTCCTCCCCAGGGCGTGAGTTCTGACCCCAG CTCCTCCCCCCATCCCCACTCCAGCCCCCTCTCCAGCTTGCTCCACCCTC TCTACCGCCCACCGGGACTGGGCATTGTCTGCCAGTCCGGGTTTCTTCCT GGGATTTGGGATGCAGAGAGGATGGGTTTGCTTGGGCGGGGGGGTGGAGA GTGAAGGGGGGAAGCAGGATCTTTGTAGAGGGAGGGACCTACAGTTACCT GGACTTCTTTCCTCTGTCTCCCCTCTTGGTACCCTTGACTGGGGCTCTTG AGGGTAATGGGTGAAGCCAAATCTGCCATGGCTCAGTTCCCAGCTCAGCT CTGTGACCTTGGGAAAGTTCCTTTAGCTCGTGGAATCTCAAGGCTCAAGG TTCCTCTTCTGCAAAATGGGGAATGATAACACCTGCCTCCTCTGGAGTCT TGGGGACTCAGTGTTCTGAGGAACGTGGCTGTAGGTCAGAGTGGCACAGA GTAGGGTCCAATGAAGCATGGCGTCCACAGTAGCTTTCCTGACTGGACTA ACCTTTCCGGACACAACAGCAGGGCAGGGGTGGGGCCTGGGGAGAAAGGA CACCTCTAACCCTGATCCTAACATCCCGATGGCCTCTAAGGCTGCCTGCA CACTCATCCAGGTGCAAGCCCTCCAAGGTGTGGTGTGATGAACCAGTGAC TCCTGGAGCCAGGTCAGCGCATCCTCTTCCCGCAGGGCTGTAAGCTGCAG GACTGAGAGGCAGGTTGACCAGGTCCTGGGCTGGATGATGGGGTGAGAGT AAGGGGTCAGTTTTGATACATGCCCAACTTTTCTCTCTAGCCCTAAGACA TCCTGGGCAAATTGCTTACCTCAGTTCCCCTGATCCTCACCCTAACCCTA ACACCAGCTCAAGAGAAAATAGGGATATTGATGGCCATCCAGAAGGGCTG CTGTGTTCCATACACAGCAATATTTCTCGAATGTTTGTGACAGCGGTCCA AGGAATAAGTTAATTTTACATTATCACTCTGGATACCTGTACAAAACTCC ACCTTATCCTTACTATATGAATGTGCTAGGGTTGTTTTTTTGTTTTGTTT TTTTTTTTTTTTTTTGAGACAGAGTTTCGCTCTTGTTGCCCAGGCTGGAG TACAATGGCGCGATCTTGGCTCACCGCAACCTCCGCTTCCCAGGTTCAAG CGATTCACCTGCCTCAGCCTTCCCGAGTAGCTGGGATTACAGGCATGCGC CACCATGCCCGGCTAATTTTGTGTTTTTAGTAGAGACAGGGTTTCTCCAT GTTGGTCAGGCTGGTACCAAACTCCCGACCTCAGGTGATCCACCTGCCTT GGCCTCCCAAAGTGCTGCAATTACAGGCATGAGCCACCGCACCCAGCCGT GCTAGGGTCTTTTTCTGTTCAATTCCTTTCTCTCTCTTGCTCTCTTTCTT TCTTTCAATGGAGTCTTACTCTGTCACCCAGGCTGGAGTGCAGTGGCAAG ATCTCAGCTCACTGCAACCTCTGCCCTCTGAGTTCAAGCAATTCTCCTGC CTCAGCCTCCCGAGTAGCTGGGATTACAGGTGCCTGCCACCACACCTAGT TAATTTTTGTACTTTTAGTAGAGATGGGGTTTTGTCATGTTGGCCAGGCT GGTCTCGAACTCCTGACCTCGTGATCTGCCTGTCTTGGCCTCCCAAAGTG CTGGGATTACAGGCATGAGCCGCCATACTCGGCCAACTTTGTATTACTTT CTTAAAGAGAGTTTCCCAAATTATATAAGCTTCAGGCCCCACAAAACCTA GATCTGCCCCAGTATAACTAAATCTGGGACCATTTATTGAGCAATTATTA TGTGCCAAGTATTGCGCTGAGTGCTTCCAGAGCATTATCTCCTTTAACCC CAGCATAGTATGTCAGATGCTGTTTTACAGATGAGCCAACTGAGACCAGA GATGCTCAGTCACTTGCCCAAGGTGACATGACTGATATGGAATAGAGTCA AGATTTTTTTTTTTTTTTTTGACACGGAGTCTCACTCTGTCTCCCAGGCT GGAGTGCAGAGGCGCAATCTCAGCTCACTGCAAGCTCTGCCTCCCAGGTT CACGCCATTCTCCTGCCTCAGCCTCCTGAGTAGCTGGGACTACAGGCACC CGCCACCACACCTGGCTAATTTTTTGTATTTTTAGCAGAGACAGGGTTTC ACCGTGTTAGCCAGGATGGTCTCGATCTCCTGACCTCGTGATCTGCCTGC CTCGGCCTCCCAAAGTGCTGGAATTACAGGTGTGAGCCACCGCGACTGGC CAGATTCAAGATTTGAACCCAGGTCCTCTTGGTCCCAGAGGCCCCTGTTT CTCAACTCCCTAGGATGGCATAGCAACCTGTCCCACAAGAGGTGCCTGCT TTAAGTGTGCTCAGCACATGGAAGCAAGTTTAGAAATGCAAGTGTATACC TGTAAAGAGGTGTGGGAGATGGGGGGGAGGGAAGAGAGAAAGAGATGCTG GTGTCCTTCATTCTCCAGTCCCTGATAGGTGCCTTTGATCCCTTCTTGAC CAGTATAGCTGCATTCTTGGCTGGGGCATTCCAACTAGAACTGCCAAATT TAGCACATAAAAATAAGGAGGCCCAGTTAAATTTGAATTTCAGATAAACA ATGAATAATTTGTTAGTATAAATATGTCCCATGCAATATCTTGTTGAAAT TAAAAAAAAAAAAAAAAGTCTTCCTTCCATCCCCACCCCTACCACTAGGC CTAAGGAATAGGGTCAGGGGCTCCAAATAGAATGTGGTTGAGAAGTGGAA TTAAGCAGGCTAATAGAAGGCAAGGGGCAAAGAAGAAACCTTGAATGCAT TGGGTGCTGGGTGCCTCCTTAAATAAGCAAGAAGGGTGCATTTTGAAGAA TTGAGATAGAAGTCTTTTTGGGCTGGGTGCAGTTGCTCGTGGTTGTAATT CCAGCACTTTGGGAGGCTGAGGCGGGAGGATCACCTGAGGTTGGGAGTTC AAGACCAGCCTCACCAACGTGGAGAAACCCTGTCTTTACTAAAAATACAA AAAATTAGCTGGTCATGGTGGCACATGCCTGTAATCCCAGCTGCTCGGGA GGCTGAGGCAGGAGAATCACTTGAACCAGGGAGGCAGAGGTTGTGGTGAG CAGAGATCGCGCCATTGCTCTCCAGCCTGGGCAACAAGAGCAAAAGTTCG TTTAAAAAAAAAAAAAAGTCCTTTCGATGTGACTGTCTCCTCCCAAATTT GTAGACCCTCTTAAGATCATGCTTTTCAGATACTTCAAAGATTCCAGAAG AT ATG

FGFR1

FGFR1 (fibroblast growth factor receptor 1) is a 100-135 kDa glycoprotein receptor tyrosine kinase specific for the fibroblast growth factor family. The FGFR1 receptor has an extracellular, transmembrane, and intracellular domain. The extracellular domain includes a single peptide and two or three Ig-like domains. The intracellular domain includes two tyrosine kinase subdomains. Stimulation of the FGFR1 receptor eventually has an effect on mitogenesis and differentiation. Specifically, FGFR1 has been associated with various diseases including Pfeiffer syndrome, various cancers, Kallmann syndrome, and osteoglyphic dysplasia.

Protein: FGFR1 Gene: FGFR1 (Homo sapiens, chromosome 8, 38411138-38468834 [NCBI Reference Sequence: NC_(—)000008.11]; start site location: 38314964; strand: negative)

Gene Identification GeneID 2260 HGNC 3688 HPRD 00634 MIM 136350

Targeted Sequences Relative upstream location Se- to gene quence Design start ID No: ID Sequence (5′-3′) site 13484 CGAGCCAGGCAGGGCCCCTCGCAAGTG 1850 13522 GACGGATATGAGTCCAGAAGTTGCG 1472 13535 TAGCTGCGTGCAGTGGCGCGCGCCTGT 4910 13561 CCGCCTCGCCAGCTCCCGAGCGCGAGTT 10239 13655 CGCCTCCTCCCAGGTGTGGGCTGGCTGCA 3067 GACCG

Target Shift Sequences Relative upstream location Sequence to gene ID start No: Sequence (5′-3′) site 13484 CGAGCCAGGCAGGGCCCCTCGCAAGTG 1850 13485 GAGCCAGGCAGGGCCCCTCG 1851 13486 AGCCAGGCAGGGCCCCTCGC 1852 13487 GCCAGGCAGGGCCCCTCGCA 1853 13488 CCAGGCAGGGCCCCTCGCAA 1854 13489 CAGGCAGGGCCCCTCGCAAG 1855 13490 AGGCAGGGCCCCTCGCAAGT 1856 13491 GGCAGGGCCCCTCGCAAGTG 1857 13492 GCAGGGCCCCTCGCAAGTGA 1858 13493 CAGGGCCCCTCGCAAGTGAG 1859 13494 AGGGCCCCTCGCAAGTGAGT 1860 13495 GGGCCCCTCGCAAGTGAGTC 1861 13496 GGCCCCTCGCAAGTGAGTCA 1862 13497 GCCCCTCGCAAGTGAGTCAG 1863 13498 CCCCTCGCAAGTGAGTCAGT 1864 13499 CCCTCGCAAGTGAGTCAGTG 1865 13500 CCTCGCAAGTGAGTCAGTGC 1866 13501 CTCGCAAGTGAGTCAGTGCT 1867 13502 TCGCAAGTGAGTCAGTGCTG 1868 13503 CGCAAGTGAGTCAGTGCTGG 1869 13504 CCGAGCCAGGCAGGGCCCCT 1849 13505 CCCGAGCCAGGCAGGGCCCC 1848 13506 CCCCGAGCCAGGCAGGGCCC 1847 13507 CCCCCGAGCCAGGCAGGGCC 1846 13508 TCCCCCGAGCCAGGCAGGGC 1845 13509 CTCCCCCGAGCCAGGCAGGG 1844 13510 CCTCCCCCGAGCCAGGCAGG 1843 13511 GCCTCCCCCGAGCCAGGCAG 1842 13512 TGCCTCCCCCGAGCCAGGCA 1841 13513 CTGCCTCCCCCGAGCCAGGC 1840 13514 CCTGCCTCCCCCGAGCCAGG 1839 13515 CCCTGCCTCCCCCGAGCCAG 1838 13516 GCCCTGCCTCCCCCGAGCCA 1837 13517 AGCCCTGCCTCCCCCGAGCC 1836 13518 CAGCCCTGCCTCCCCCGAGC 1835 13519 TCAGCCCTGCCTCCCCCGAG 1834 13520 TTCAGCCCTGCCTCCCCCGA 1833 13521 CTTCAGCCCTGCCTCCCCCG 1832 13522 GACGGATATGAGTCCAGAAGTTGCG 1472 13523 ACGGATATGAGTCCAGAAGT 1473 13524 CGGATATGAGTCCAGAAGTT 1474 13525 TGACGGATATGAGTCCAGAA 1471 13526 CTGACGGATATGAGTCCAGA 1470 13527 TCTGACGGATATGAGTCCAG 1469 13528 GTCTGACGGATATGAGTCCA 1468 13529 TGTCTGACGGATATGAGTCC 1467 13530 ATGTCTGACGGATATGAGTC 1466 13531 GATGTCTGACGGATATGAGT 1465 13532 TGATGTCTGACGGATATGAG 1464 13533 GTGATGTCTGACGGATATGA 1463 13534 AGTGATGTCTGACGGATATG 1462 13535 TAGCTGCGTGCAGTGGCGCGCGCCTGT 4910 13536 AGCTGCGTGCAGTGGCGCGC 4911 13537 GCTGCGTGCAGTGGCGCGCG 4912 13538 CTGCGTGCAGTGGCGCGCGC 4913 13539 TGCGTGCAGTGGCGCGCGCC 4914 13540 GCGTGCAGTGGCGCGCGCCT 4915 13541 CGTGCAGTGGCGCGCGCCTG 4916 13542 GTGCAGTGGCGCGCGCCTGT 4917 13543 TGCAGTGGCGCGCGCCTGTA 4918 13544 GCAGTGGCGCGCGCCTGTAG 4919 13545 CAGTGGCGCGCGCCTGTAGT 4920 13546 AGTGGCGCGCGCCTGTAGTC 4921 13547 GTGGCGCGCGCCTGTAGTCC 4922 13548 TGGCGCGCGCCTGTAGTCCC 4923 13549 GGCGCGCGCCTGTAGTCCCA 4924 13550 GCGCGCGCCTGTAGTCCCAG 4925 13551 CGCGCGCCTGTAGTCCCAGC 4926 13552 GCGCGCCTGTAGTCCCAGCT 4927 13553 CGCGCCTGTAGTCCCAGCTA 4928 13554 GCGCCTGTAGTCCCAGCTAC 4929 13555 CGCCTGTAGTCCCAGCTACT 4930 13556 TTAGCTGCGTGCAGTGGCGC 4909 13557 ATTAGCTGCGTGCAGTGGCG 4908 13558 AATTAGCTGCGTGCAGTGGC 4907 13559 AAATTAGCTGCGTGCAGTGG 4906 13560 AAAATTAGCTGCGTGCAGTG 4905 13561 CCGCCTCGCCAGCTCCCGAGCGCGAGTT 10239 13562 CGCCTCGCCAGCTCCCGAGC 10240 13563 GCCTCGCCAGCTCCCGAGCG 10241 13564 CCTCGCCAGCTCCCGAGCGC 10242 13565 CTCGCCAGCTCCCGAGCGCG 10243 13566 TCGCCAGCTCCCGAGCGCGA 10244 13567 CGCCAGCTCCCGAGCGCGAG 10245 13568 GCCAGCTCCCGAGCGCGAGT 10246 13569 CCAGCTCCCGAGCGCGAGTT 10247 13570 CAGCTCCCGAGCGCGAGTTG 10248 13571 AGCTCCCGAGCGCGAGTTGG 10249 13572 GCTCCCGAGCGCGAGTTGGA 10250 13573 CTCCCGAGCGCGAGTTGGAG 10251 13574 TCCCGAGCGCGAGTTGGAGG 10252 13575 CCCGAGCGCGAGTTGGAGGA 10253 13576 GCCGCCTCGCCAGCTCCCGA 10238 13577 CGCCGCCTCGCCAGCTCCCG 10237 13578 CCGCCGCCTCGCCAGCTCCC 10236 13579 GCCGCCGCCTCGCCAGCTCC 10235 13580 CGCCGCCGCCTCGCCAGCTC 10234 13581 CCGCCGCCGCCTCGCCAGCT 10233 13582 GCCGCCGCCGCCTCGCCAGC 10232 13583 AGCCGCCGCCGCCTCGCCAG 10231 13584 GAGCCGCCGCCGCCTCGCCA 10230 13585 GGAGCCGCCGCCGCCTCGCC 10229 13586 AGGAGCCGCCGCCGCCTCGC 10228 13587 GAGGAGCCGCCGCCGCCTCG 10227 13588 TGAGGAGCCGCCGCCGCCTC 10226 13589 CTGAGGAGCCGCCGCCGCCT 10225 13590 ACTGAGGAGCCGCCGCCGCC 10224 13591 CACTGAGGAGCCGCCGCCGC 10223 13592 TCACTGAGGAGCCGCCGCCG 10222 13593 CTCACTGAGGAGCCGCCGCC 10221 13594 ACTCACTGAGGAGCCGCCGC 10220 13595 GACTCACTGAGGAGCCGCCG 10219 13596 GGACTCACTGAGGAGCCGCC 10218 13597 GGGACTCACTGAGGAGCCGC 10217 13598 CGGGACTCACTGAGGAGCCG 10216 13599 CCGGGACTCACTGAGGAGCC 10215 13600 CCCGGGACTCACTGAGGAGC 10214 13601 TCCCGGGACTCACTGAGGAG 10213 13602 CTCCCGGGACTCACTGAGGA 10212 13603 CCTCCCGGGACTCACTGAGG 10211 13604 CCCTCCCGGGACTCACTGAG 10210 13605 TCCCTCCCGGGACTCACTGA 10209 13606 GTCCCTCCCGGGACTCACTG 10208 13607 TGTCCCTCCCGGGACTCACT 10207 13608 CTGTCCCTCCCGGGACTCAC 10206 13609 CCTGTCCCTCCCGGGACTCA 10205 13610 GCCTGTCCCTCCCGGGACTC 10204 13611 GGCCTGTCCCTCCCGGGACT 10203 13612 GGGCCTGTCCCTCCCGGGAC 10202 13613 CGGGCCTGTCCCTCCCGGGA 10201 13614 CCGGGCCTGTCCCTCCCGGG 10200 13615 CCCGGGCCTGTCCCTCCCGG 10199 13616 CCCCGGGCCTGTCCCTCCCG 10198 13617 GCCCCGGGCCTGTCCCTCCC 10197 13618 CGCCCCGGGCCTGTCCCTCC 10196 13619 TCGCCCCGGGCCTGTCCCTC 10195 13620 TTCGCCCCGGGCCTGTCCCT 10194 13621 CTTCGCCCCGGGCCTGTCCC 10193 13622 CCTTCGCCCCGGGCCTGTCC 10192 13623 GCCTTCGCCCCGGGCCTGTC 10191 13624 CGCCTTCGCCCCGGGCCTGT 10190 13625 CCGCCTTCGCCCCGGGCCTG 10189 13626 GCCGCCTTCGCCCCGGGCCT 10188 13627 CGCCGCCTTCGCCCCGGGCC 10187 13628 TCGCCGCCTTCGCCCCGGGC 10186 13629 CTCGCCGCCTTCGCCCCGGG 10185 13630 CCTCGCCGCCTTCGCCCCGG 10184 13631 GCCTCGCCGCCTTCGCCCCG 10183 13632 GGCCTCGCCGCCTTCGCCCC 10182 13633 GGGCCTCGCCGCCTTCGCCC 10181 13634 CGGGCCTCGCCGCCTTCGCC 10180 13635 GCGGGCCTCGCCGCCTTCGC 10179 13636 CGCGGGCCTCGCCGCCTTCG 10178 13637 CCGCGGGCCTCGCCGCCTTC 10177 13638 ACCGCGGGCCTCGCCGCCTT 10176 13639 AACCGCGGGCCTCGCCGCCT 10175 13640 AAACCGCGGGCCTCGCCGCC 10174 13641 GAAACCGCGGGCCTCGCCGC 10173 13642 GGAAACCGCGGGCCTCGCCG 10172 13643 AGGAAACCGCGGGCCTCGCC 10171 13644 CAGGAAACCGCGGGCCTCGC 10170 13645 CCAGGAAACCGCGGGCCTCG 10169 13646 TCCAGGAAACCGCGGGCCTC 10168 13647 GTCCAGGAAACCGCGGGCCT 10167 13648 AGTCCAGGAAACCGCGGGCC 10166 13649 CAGTCCAGGAAACCGCGGGC 10165 13650 CCAGTCCAGGAAACCGCGGG 10164 13651 CCCAGTCCAGGAAACCGCGG 10163 13652 CCCCAGTCCAGGAAACCGCG 10162 13653 TCCCCAGTCCAGGAAACCGC 10161 13654 CTCCCCAGTCCAGGAAACCG 10160 13655 CGCCTCCTCCCAGGTGTGGGCTGGCTGCAGACCG 3067 13656 CCGCCTCCTCCCAGGTGTGG 3066 13657 GCCGCCTCCTCCCAGGTGTG 3065 13658 TGCCGCCTCCTCCCAGGTGT 3064 13659 CTGCCGCCTCCTCCCAGGTG 3063 13660 CCTGCCGCCTCCTCCCAGGT 3062 13661 GCCTGCCGCCTCCTCCCAGG 3061 13662 AGCCTGCCGCCTCCTCCCAG 3060 13663 AAGCCTGCCGCCTCCTCCCA 3059 13664 AAAGCCTGCCGCCTCCTCCC 3058 13665 AAAAGCCTGCCGCCTCCTCC 3057 13666 GAAAAGCCTGCCGCCTCCTC 3056 13667 AGAAAAGCCTGCCGCCTCCT 3055 13668 CAGAAAAGCCTGCCGCCTCC 3054 13669 CCAGAAAAGCCTGCCGCCTC 3053 13670 CCCAGAAAAGCCTGCCGCCT 3052 13671 CCCCAGAAAAGCCTGCCGCC 3051 13672 TCCCCAGAAAAGCCTGCCGC 3050 13673 GTCCCCAGAAAAGCCTGCCG 3049

Hot Zones (Relative upstream location to gene start site) 1350-1500 1750-1900 2500-5500 10150-10300

Examples

Genetic Code (5′ Upstream Region) (SEQ ID NO: 13679) AGCTGGCAGGGCGAAGGGCCGACAAATCCTCCCTGACCCTCCCAGCTCTT TGTTATCTCAGAGGGAAGGTTACATTTCTGTATGGGAGGCAAGGTGCCAG GAGGCCTCGGGCAGAACAGAGACAGGCAGAGCTGCTGTCTGACCCCTGTT GCCTGGAGCAGCTCAGGGCTGCCCTAGGGACACTCTCCCTCCACTGGCCT GGGGCCCTTCCAGAAATGGGAGGGCTACATTTCAGAAAGAGGGCGAGTAG AGGAGTGGGACAGAAAAGGAGCGAGGTGGGCTGGAAGGATAAAAGCAGCC AACTCTCAATTATTCAGAAACCTGTCTGCAGTGTGTGGACAGCCCATGCC TTTGCTGAGTTTCTCACCTTCTCTGTTCAGCTGCCATCAGCTCTTTCCCT GAGAAGTGGAGGAGGGACCCTGGCAAGTTGGCCACTTGCTTTCATTTTGG CTTCTTGATAAATCTATAGAGGATTTTTCAGCAGCAGGCCCATGTCCCTC AACCCCAAACAAGCATTTAGATCATTATCTTTCTGTTTAAATCAAGAACG CATTATTTAGCCTTTTATTTGGGGTTCAAGATACTCCTACAATGGTTCTA AATCATAAGAAAAAGGGGCTTGATTTAAAACCCCTTGTTTTGGGCCAGGA ATGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGTGGGCA GATTACCTGAGGTCAGGAGTTCGAGACCAGCCTGGCCAACATGGTGAAAC CCTGTCTCTACTAAAAATACAAAAATTAGCTGGGCATGGTGGCAGGTGTC TGTAATCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATCGCTTGAGCCCA GAAGGCAGAGGTGAGCTGAGTGAGCTGAGATCGTGCCATTGCACTCCAGC CTGGGCAAAAAGAGCAAGACTCCATCTCAGGAAAAAAAAAAAAAAAGAAA ACAAAAAAAACCCTTTTTTGAGAAGAATTACGGAGCAAAGTAGAAAAATA GTAGCTGGGTGTTAACATTAAATGCTGGATTTTTTTCATGGCTTGTCTTC CCAATCATATTCCCTCAAATTGTGTTTCCTCCTCTGGTAACCCAGGTTGG TTATGCTTAGCAAGTCCATGAACAATAAATATACATGGAAAACCTCCTGT GTAGAATTGGTCAGACACCTAGATAAGATCCTTGCCCTAAAGCAGTTTAG AAACCAGTTAGAAAGAAAGCAGAGTAAGGAAAACCACTAACAAAGCACGG TATCAACTCAGTGGATAGTCAGCAAGTGAGCAGGGGGTCCAGGGACTGAC AAAGCTGGGATGGGCAGGGAAGGCCTCTTGGGGGTAGGGTGTGAGTATGG CCTTCTTACAAGCGTGTGATGTGTAGTAATTAAAATGCAGGAGGCCTAAT GGGTGGGCAGCTTACATAGGAGTATAAACCAAGCTTGACCAGGAGCTGAA AGGTTAAATGGTGGCTCTTAGGGGAAAACCCTATAAACAGTGGCTGAAGT TCATTTATTCAACAAAGATATGAGTTCTTGTTTCTCATTTTTTGTTTTGT ATTATTTTGTTTTGAGACAGGGTCTTACTCTGTCGCCCAGGCTGGAGTGT AGTGGCTGGATCATAGCTCACTGCAGCCTCAAACTCCTGGGCTCAAGCCA TCCTCCTTCTTCAGCCTCCACCTCCAGCTAATTTTTAAAAATATTTTGTA GAGACAAGGGCTCACTTTGTTTCCCAGGCTGGTCTTGAACTTCTGGCTTC AAGTGATCCTCCCGCTTCGGCCACCCAAAGTGCTGGGATTACAGGCGTGA GCTGTAATTTAGTTGTTTATTTACTCATTTGTTCAACAAATACTTATTGA ATATTTGCTCTTTGGCCAGTCAAGGGATTTCATGAGTGTCTACTATGTGA ATAACACTGTGTTGGCCACTAGTCTGTCACCTACTGGTGGATTAGAAAAA TAGCGCGAGGACCATTTTTTCTTTTCTTTTCTTTTTTTTTTTGAGACGGA GTCTTGCTCTGTTGCCAGGCTGGAGTGCAGTGGCACAATCTCGGCTCACT GCAACCTCCGCCTCCCGGGTTCAAGCGATTCCTCTGCCGCAGCCTCCCCA GTAGCTGGGATTACAGGCAAGCGCCACCATGCCTGGCTAATTTTTTTGTA TTTTAGTAGAGACGGGGTTTCACCTTGTTGGCAAGGATAGTCTCGATCTC CCGACCTCGTGATCCACCCGCCTCGGCCTCCCAAAGTGCTGGGATTACAG GCATAAGCCACCGCACCCGGCCAACTCTTTTCTTAAATTAGCCAGGGAGG CGTGGGTGGGTTGGGTGAGGAGTTGGGTGGGGGGATCTCATTCAGTATTC AAACTTCTACAAGTTTCGGGGTTGAGGTGGGTGATGGTAAGGGAACAGGC CCTGCCACTACCTTTCATAGTGACTTCCATTTGTGTAATATTTTTGGTCC ACTGAGAGCTATTATTTTATTTGATTCTTATGACCATCTTGTGAAGGAGT ATCAACAGATACCCCGTTTTGATTTTATCAGATGCATGATTTGTCCTACA TCAAACTTCATAAATGATGGACAGAATGGAGGAATCCTTCAGACCAAGTG CTGCCTACTTCCCACCCCAATGGTGGCCTCAGCCTGGGCTCACATCACAC GCCCCAAGGAGCCTTGGAAAAAATAAAGGCTCTTGGCTCCTTCCTGGGAC AGCGTGATTCCTCATGTCTGAGCAGGCCCATGAACTTGTATTTTTCAGAC GTTCCCTAGGACCCGTGTCCATCTGGATTAGGGAACCACTACATTATACC ACTTCGCGGGAAGACTCAGGGGGAAGCATTTTAGCCACTTTCCTGTGTTC CACAGTACTGGAGGGTGTTCTGAGTGGGCTGTGATTAATTTCCAAACCAA CCACACGTCTCCCCTCAACTCCCACTGCTTACTCTTTGCTTCCTAGACAT TCACTGCAGGCTGGAGACTTCTGGAAGCCAACAGCATCGCTGTAGAATTT ACAGGGTCCAGTTCCCGGTGGACCACAAAACCTAAATTATGTGGCTGGGG AAAGCTGAAATCCAAGGGAAGGGTTTGAGGAGGGGCTGACCTTATAATAA AACCGGCTTGTATTTACTAAGTGTTAACTATGCGCTAGGCCCTCGTTGAC GCCTCAACTCTATGTGAAAAGCACTATTATCCCCCATTTACAGATGGGAA AACAGAGATTTAGAGCGCGAAAATCATTTCCCCAAGGCGCACAGACTCCA AAGCCCACGCTACCAGGTACAACCTCAAGGCTGCGGCGTCTCTTCACCTG CCCCCTAGCCCCCAAACCGCTGCTATGTCTAGGGCCTGACATTCCGGCGC CCTCTGGGACGTGCTCAGATGCAGGGGCGCAAACGCCAAAGGAGACCAGG CTGTAGGAAGAGAAGGGCAGAGCGCCGGACAGCTCGGCCCGCTCCCCGTC CTTTGGGGCCGCGGCTGGGGAACTACAAGGCCCAGCAGGCAGCTGCAGGG GGCGGAGGCGGAGGAGGGACCAGCGCGGGTGGGAGTGAGAGAGCGAGCCC TCGCGCCCCGCCGGCGCATAGCGCTCGGAGCGCTCTTGCGGCCACAGGCG CGGCGTCCTCGGCGGCGGGCGGCAGCTAGCGGGAGCCGGGACGCCGGTGC AGCCGCAGCGCGCGGAGGAACCCGGGTGTGCCGGGAGCTGGGCGGCCACG TCCGGACGGGACCGAGACCCCTCGTAGCGCATTGCGGCGACCTCGCCTTC CCCGGCCGCGAGCGCGCCGCTGCTTGAAAAGCCGCGGAACCCAAGGACTT TTCTCCGGTCCGAGCTCGGGGCGCCCCGCAGGGCGCACGGTACCCGTGCT GCAGTCGGGCACGCCGCGGCGCCGGGGCCTCCGCAGGGCGATGGAGCCCG GTCTGCAAGGAAAGTGAGGCGCCGCCGCTGCGTTCTGGAGGAGGGGGGCA CAAGGTCTGGAGACCCCGGGTGGCGGACGGGAGCCCTCCCCCCGCCCCGC CTCCGGGGCACCAGCTCCGGCTCCATTGTTCCCGCCCGGGCTGGAGGCGC CGAGCACCGAGCGCCGCCGGGAGTCGAGCGCCGGCCGCGGAGCTCTTGCG ACCCCGCCAGGACCCGAACAGAGCCCGGGGGCGGCGGGCCGGAGCCGGGG ACGCGGGCACACGCCCGCTCGCACAAGCCACGGCGGACTCTCCCGAGGCG GAACCTCCACGCCGAGCGAGGTAAGAGCCGCGGCGCCCCCGGATCTGGGG CGGGCTTGGCGTCCCGAGCGGCCCCCGGCGCCGGAGCCTCCCGGCTGCGC GCTTTGCCCGCCGCAGCCCAGCCGGGGCCGGCGCCTCCCTCCGCTCGCCG CCCGCCCCTTTCACCTCCTGGCTCCCTCCCGGGCGATCCGCGCCCCTTGG GTCTCCCCTCCCTTCCCTCCGTCCGCGTCTCCTGCGCCCCCTCCCTGCGC TCGTCCCGCCGCTCTTCCCGCCGCCCAACTTTTCCTCCAACTCGCGCTCG GGAGCTGGCGAGGCGGCGGCGGCTCCTCAGTGAGTCCCGGGAGGGACAGG CCCGGGGCGAAGGCGGCGAGGCCCGCGGTTTCCTGGACTGGGGAGGAGGG CGGGAGTGGGCGGCGAGGTGGGATGCGTTGTGTGTGTTATGTGTGTGTGT TGCATTCCACTCCATGTCTTTTTGGTCCCCTTTTGGGGATTCACCCCCAA TTCAGCAGGTAGCTTTGGGCTCAACGCTAAAAATCCGGGGCATTCCTAAG TCCTTTTCCACCCCCGGGAAAGCCTGGGGTGCGGGTTGGGGTCGGATGGG GTGGGAGATGAACTGCGGAGGACGTGGAGGGCTAGGTTAGCTTCTCTTGG AATAGGTTTTAAGGAGGTGTCGTCACCAAATGGCTGAATCTGCTTGAGCT GAGAGCGAAAAACGACTCCCCTTTCCAGAAGGGGTGATCTTATGACTTGG ACGGTCTCTGAAAGGGTCGGAAGTTTGGGGAACGGGAGGACAACCCACGG TCGTTAAGCCGAGGTGTGGGATGGGGGCGGAAGGACCGTTCGGTCCCAAT CTGGTTCCTAGAGGTGGGGGAAGGGATGAGGGTTTTTGTCCGGTGTGGTT CACTCGGCAGCGATGCGTATGCTTCTCTGGCCCAGACCCCTCTGCACCTC GCTTCCCCTACCGTTATGTTTGGGGTTGGGAGAAAAGTGAGGCTACGACC CATGTTTGCGGAGGAATTTTATGGACCTTGTAGATGGGGGTTCATATAGA ACACACACCCCCTATGAGGCAGCCAGACACTTTTTTGGTGGTGGTGGGGG GGGGGTGGGGTGTGAAGCCTGTTTCTTGTTCTGAGCCCAGAAGCTATCAA CCCTTTTGAAAAACATTACCACGGTGCCTTTCTCCCCCAGCACTCCCCCA CCCCCAATTTCCAGATGTAGCAGCCGCATCTGGTTCCGTTTCACCCCACA CGGGTACACCGCAGCCGCATTATTAACTTCCCTCTTCCTCCCCTCCCCCT CCCCCAAATTAAAACTCAGATTCTTCAGCCTGTCTTGACCACCTCCCTCC TTAACATTTCTGGAGACTTGGAGATGCGGCGTTGAGATTCGGGGGAGAAA AGAAAGTTCCCTTGGATCCCGAGTTATTTAAGATCTCACCAAGTTATTCG CCGCCGCTGGTGGGTGGCGGCGGTCCGGGTGCTTTCTGGATTGCGCAGTA AAGAGGCATCTTGGGAGATGGGGCCAAGGTTTTAGGGGGTGCCACTCGCG AACGGTTCATCCGCTAGACTAGGGGGGCTCTTTGGCTGTGCGTCTGGCCA GAACTGGCCTTGACGATGGAAGTTTCTGGAACCAAAGCGTTGCTTTCTCT CCCTTGTGTTATAGCTGGAGCTGCGGGAGCGCCTGCCCTGCCCGGAGCCC GCGGTCCCCTCTCGGCTGCCCCGCGGTGGCGTCACGCGCCCCTCCCGGAG CAAGCCCGGTGCGCAGGGCCGGGGGCGTGGGCGGCTGCTGCCAGAGGCGC TCTCTGTGTGTTTTTAAGGACTGATTTGGGCCGCATCCCCCGGAAACTAA AGTGGGGTGTTTTACCGTTTAAATAACGGCTACAGGTTTGAAAGCGGGGT TGGATTTTCGAGTTGTGTTTGGTAATAGTCTTTGAGGCAGGAAAGCGCCT TGTGGTCCAAAGTTGCCGGGAGGGTGGGGAGAGTCGGTGTCTTACCCGCT TCTTTCCAGCCTCTTTCAAATTGAAAACACTTCTCTGGTTTCCTTCTTTG GGCGGTAGTTTTGGAGGCTGTAATGAAATCGCACTTTCTCTAGACGTGGT AATTAAGGTGACTGTTTCCTCCGCAGATGTGCCCTACCCTTTGCACCTCC GGACCAGCGCTTTTTTTGGAATACTATCTAGCCTTGAGACTGTTTAGCAG AAAGTGGCCATTTTCCTCCCTTGGCCCGGGCTCCCGGTTTCCTCCCTGAG GCTTGTTTAAAAGCGAAGTAGCAGGGCCCCGTGGGACGCGCCTTGGTCTG GGTAATCACCCCCACGCCCGGGTCATCCACCTTCCTCTCGGTGACCGAGG TTCAGCAGCCTCTGCTATTGCCGGCCGTCTTTGCCGATGGCCTGCCTCCC TAATGACTTGTTTACATATCCTACCCCCAGTGGGTTAGGAGAAGCTCCGG GGCTGCCCCGACCCTCCGAGTGCAGGGTGTTTGGGGACCGGGAGGCTGCT GGGGCCTGACTCCAGCTGGGAGGGTTATGAACTGCATCAGTGACGAGCTG CTTGAAATATCTGTTGCATTTACTCTTAGTCATAGCTGAGTGTCAGCTTT TTAATGAGGTTCATCCAGATTGAGAGCCACTTGGACTGCGTACTTCACTG CCTGCTTTTCCAAACATGCCTGCAGAAATGCTCATTTTCGAGGTATTTTT CCCAATGGGAATTCAGGCCAGAGTGGGCACCACTTGAACAATCTTAGGGT GCTTCTTTTCCTTGGCCTCTGGCCATGGAGGGTGTTAGACAGTTCCATTA GGTGGCCCTTTGATAGCAAGGGAAGCAAAGGCTCAGGAAGAAATGGAGAA GCGTCCCCCACTCCCTAGGGGCAGAGGATTAGATACATCGGTGCATCCCT CAGGCTGGGCTAGCTTTATTCCTGGTGGACTCCAGAGGGCAAGAAAATTG AATTGAACACTGGGTAGGCAGATTCAAGCCTTAGAGACCAAGGAAAATCC ATGGGTTTTGCTTTTAGTGGTGTGCTCTTTGTTTTCAGTATTGACCTGAA ACAAGACTCCTAAAATGAGAGATTTGCTGGTATGAACTTGGGGGTTTAGC AGCCGGCTTCTACAAAGGCTTTTTTCTTGCCTTCGTTTCTAAAGTGTCTT TCGTCAAAATGGCTGTTAGTTATAGAACATCCTAGCAAAGTTTGAGCCTG TTGCTGCTGGAGGAAAAGGAGTTAGAATTGATTCAAATGTCTTATTCTGA AAGGGCCTCACATCACTTGATAGTTTAATTTCCTCCTGGGAAATTTGTGT CTTACATTTGTCTTCCCCAGAGCTTTGTAAAAGGCCTGAACGCACCAGGG ACTAGTGGGAGCCCAGATGCAGAGCTTTAGAGAAGATTCTGGTGTTTCCA GAGAGGATGAAATGTCAGACTTGGGCTAGGATATTTGTTTTTCCTCCTAA GGTTGCATCTACTTTAAACAGAAATTCTCTCCTCGCCACCATTTATCTCT CCCCTGCAATGAAAGAAACCATGTTTAGGGCCCTCTCCCCCATTTAATAG CCCTCACATGGATGAACTATCCCAAGAATTTGGTGGGGTTCCACTCATAG TACATCCTGTCTTCAAGAGCAAGGTTTTCTAGATTATGTGCAGCAGTTCG TGTTTCACTTGTTGCTTTTTTTTTTTTTTTTTTTTTTTGAGATAGTCTCG CTCTGTCGCCCAGGCTGGAGTGCTGTGGCGCTATCTCAGGTCACTGCAAC CTCCGCCTTCCGGTTGAAGCGATTCTCCTGCCCCAGCCTCCCTAGTAGCT GGGATTGCAAGCATGCGCCACCATGTCCGGCTAATTTTTTGTGTTTTTAA TAGAGATGGTGTTTCACCATGTTGGCCAGGCTGGGCTTGAACTCCTGACC TCAAGCAATCCGCTGGCCTCGGCCTCCCAAAATGCTGGGATTACAGGTGT GAGCCATTGTGCCTGACCACTTATTGCTAATTTTTTATATGTCTCTTACT TCCAAGGACATTTAGACACTTTTTTTTTTTAAAGAGACTCAAAAAATTAG CATTTCCATTGGACCAACTAAAATTTAGCAAGCTGAGCTGAGTAACTTTC TCCATATGTTTATTAAGTACTTGCCCCCTGCCCTCTCAACATGTGAGTAG AGAATGGTCACTTTGGGGAAGAAATAAGTCTTATTCTCATCTGAAGGGAT TAATGTTTTGGTGTTACTTCCTCAATTCTGAAGAACCAAGTTGTCCAGAA ATTTTCTCAGGGTTCTTTGGACTAGAGTTTGGCTGGTTAACAAGGGGTAC TACCTAATTGCTTTTCTCTGATATTCTCAGCCTCTTTTTCTGGAGGAGTA TCTCTGTCAGTTTCTTTTCATCAGCCCTTTTTTTTCCTTCATTCACTTAC TCATTCATCCAGTTAACAAACATGTTGGCATCTCCTGTGTACATGCTAGG TGCCGAGGGTGTTAGCAAAGGTTAGGGAGGCACAGACCCTGTTCTGAAGG AGCCTGCAGTTTCGTGGGGAGAGAAGAGAATGAAGAACATAAATAACAAT CATATAATATGACCTAAGTGCTATGTGAGAGGGGCTAGTAATGTGGTTTG CAAATTTGGAGGAATGAAATTCTCCAGCTAGAAGGCCCAAGAAAGTCTTA TGGAAGAAACAGCTTCTTAAGGTGGGGTTCAGAGAAAAGGGAAGGGCTGG CCTGTTGCAGAACAAGGAATGGCATGAAGAAAGTCTTGCACAGAGGCATG GATGTTGCTTCGAGCTGTGGCGCCCTATAGAAATAGAACATGAGCAGCTG GTCACAGTGGCTCATGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGCAG GCGGATTGCTTGAGCCCATGAGATGGAGATGAGCCTGGACAACATGGTGA GACCCTGTGTCTACCAAAAAATACACAAATTAGATGAGTATGCTCGTGCT TACTGGTAGTCCCGGCTATTCAGGAGGCTGAGGTGGGAGGATCACTTGAG CCTAGGAGGCAGAGGCTGCAATAAGCTGTGATTGCACCACTGCATTCCAG CCTGGGGGACAGAGGAAGACCCTGTTTAAAAAAAAAAAAAAAAAAAAAGC CAGGCACAGTGGCTCATGCCTGTAATCCCAGCCCTTTGGGAGGCCAAGGC AGGTGGATCACCTGAGGTCAGGAGTTCAAGACCAGCCTGGCCAACATGGT GAAACCCTATTTCTACTAAAAATAAAAAAATTAGCCGGGCTTGGTGGCTC ATGTCTGTAATCCCAGCTACTTGGGAGGCAGGAGAATCGTTTGAACCCGG GAGGCGGTGGTTGAGCCAAGATTGCGCCACTGCAACTCCAGCCTGAGTGA CAGAGCAAGACTCCATCTCAAAGAAAAAAAGAAAGGAAGAAAGAAATATA ACATTATAACATGAGTTATGTATATGTTCAGATTTTCTAGAAGCCACATT GGAAATTAAGTTAAAAGAAAGAAATAGGTAAAAAAAATTTTTTTTTTTGA GACGGAGTCTCACTTTGTTGCCAGGCTGGAGTGCAGTGGCGCAATCTCGG CTCACTGCAACCTCTGCCTCCCGGGTTCAAGCAATTCTCCTGCCTCAGCC TCCTGAGTAGCTGGGACTACAGGCGCGCGCCACTGCACGCAGCTAATTTT TGTACTTTTAGTAGAGACGGGGTTTCACCATGTTGGCCAGGATGGTGTCG ACCTCTTGACCTCGTGATTTGCCCACCTCAGCCTCCCAAAGTGCTGGGAT TACAGGCGTGAGCCACCGCGCCTGGCCAATATTTGTTTTTTAATTAACTT GTTTGTTTAGATTTTATTTAATGTAACTATATTTCCAAAATATTATCATT TGAACATGTAATCAATATAGAAATTATTGATGAGATACTTTACATTTTTT TCATAACAAGTTTTTAAGATGCGGTGTATACTTTTTACTTATAGCATATC CGTTAGCACCAGCCACATTTCAAGTGTGCAGTGGCCACTGTGTGGGCCAC AGGTCTAGAATATAAGACATGAAGATGGAGAGTGAGAAATGCCTTTGGAA AGGTTGGAAGTTCCTGTCCTTCTGCTGCCAATTACCAAATCTCCTGAGAG TGCTATTAAGGAGTGACTCAAAGCACTACACAAAGAGAATTATAAATATC TTAATATTATATCTGAAATCCAAATGCATAATTCTTTACATTTGGTTGGT ACTTTAGAGAGGAGAGAATGGGCACAGTCACCCACACCACCCATTTGAGC CTCATAATCACCTGTGATGTGGCTTCCTCTAGGTGGGAAACCGAGGCTTA GAACGGTTAAGTGACTATCCCAGGGTGGCAAGATCATAAGTGGAAGGGTG TGAATTCATACTGTCTCCAGCGGACAAGAATAAAAAGACCCAGGCTGGGT GTGGTGGCTCATGCCTGTAATCCCAGCACTTTGGGAGGCCACTGTAGGTG GATCTCCTGAGCCCAGGAGTTCATTACCAGCATGGGCAACATGGTGAGAC CCCATTTTTATTAAATATACAGAAAATTAGCCCAGCTTCTCGGGAGGCTG AGGTGGGAGGATCACTTGAGTCTGGGGGATGGAGGTTGTAGTGAGTTGAG ATCGTGCCACTGCACTCTAGCTTGGGTGACAGAGCAACACTCTGTCTCAG AAAGAATAAAAAGATTTGGCCATGAATTCGTCAGCTAGTTTTCCTTACAT AATTTTTGGACAAGGAGATCTGACATTCATAGGTTTTTCTCTTAGAAGTG GGAGAGCTTCAAGGTCACGTGGTCCGTCCAGCCCCTGCTATCTCACCAGA CACTGTCCACCCTGTATGTTGGATCAGTACTCCAGTGAGAAGACAGCAGG CACTTTCACCCATGCAGCCCATTCAGTCTTCATAACCACCTGTGATGGAG GCAAGGCAAGTATTTCAGCCCCCTCTGATGAGTGGGAAACTGAGATGTGC CCCCTCTCTGCTCCCCACCGAGGACCTCTGCATGCAGGCATGAATCCCAG GAGCCTAGCTGATATTGGAGAGACGGGGCGGGGGGAACCAGCTGCAGGGT CTTGGAGGAAGCTGCTGTGTACACCTGCAAGGCTGCAGGTTACATCTATC TGTCAAGCAGTGAAGGAAGGAAGTTGTTTCTAAGGGATTGGAAAAATTCA TTAATTAGTAGAATGAGAAACTGAGGTGAAGCAGGAGGTGGCAGGGTCCC AGACAGCATGTTGGACTAGTGGCCTGTGTCACTGTGTTTTTTGCAGGCGG GTGGCATGGGGTGTATGCTGACTTCTTATTCCAGGAGTTGGTGCCAGGAG GCCAGGTTTTCTTAACATCCTTGTTTTACAGATGTCAAACTTGAGGGCCA GAGGGGTAGGAGAGGAAGAGACTTTTTGTACCTTTTTTGGGAAAGAACAA GAGGGAAGCTGGCAGATGAATTTGAAGTGCATTGACCAGGGAGCTGAGAG AGGGCGGTCTGCAGCCAGCCCACACCTGGGAGGAGGCGGCAGGCTTTTCT GGGGACAGAGTGGCCAAGTCGAAGCAAGCTTAACCATCTCAACATGACAC CACTCTTTCCCATTGGAACCTGAGAACTTGTTCAGTATTCTGACACTTAG CAAGGGACCTGGGTTTTCTTGGTCAGGTGTGCGTTTCTGGGTGACAGGCC TGCATCAGGTGTATTTTCGGGATGTAGTAAGTTGTGGAATATGGGTTTAG GGGCATCCTCTGGCAAGCACTGCTTCTATCCCAGCTCTGGGAATGTGCCC CATGCAGTGTCCTAGATGGCCCATCTGTGGTCTGCTTCCAAGGGTCTTTC TTTTAGTTAGTTAGTTTTGAGACAGAGTCTCACTCCGTCACCCAGGCTGG AGTGCAGTGATGCAATCTCGGCTCACTGCAACCTCCACCTCCCAAATTCA AGCAATTCTCATGCGTTAGCCTCCTGAGTAGCTGGGATTACAGGCGTGCA CCACCACACCCAGCTAATTTTTGTATTTTTAGTAGACGAGGAATTTCACC ATGTTGGCCAGTCTGGTCTCAACTCCCCACCTCAGGTGATACTCCCGCCT CAGCCTCCCAAAGTCCCGGGATTGTAGGTATGAGCCAACATGCCCTGGCA CAAGGGTCTATCTTTGACCAATGGAACTGCAAATCAAGCCTCTTTTGTTA CCAGAGTTACCTTGGATTTACCCTTATCTACTTGGTTTGGATAAATTGAG TTTGCATCAGATGGAGTCAGGCTTGATCAATCCCTTATTTACTTCCTCCC ACCCTGTTCTCTAATATCCAAAAACCTTGAGGCACTATTACATGCTAGCT ACATTTCCTTGAGTAAAGTACTTAACCTCTTTGAGCCTCAGTTTCTCCAT TGCATAAAAGGAATAATAAAACTTATCCCCCATAAGTTTATAGTGAGGAA TGAATTAATTCCTCACTATAGTTCTAAATTAATTCTACTTAGGGCATCCT TGGTACATAGTGGGTGTTCAGTATTCATTTCATTTTCTCTTTTCTGATTC CTTTCGTAAAAGTAGAAAAATGAAAGAGAAATGTTGACTTCTCTTTTGAT TTGAAATCATTAAAACATTTTAGTAAGCCTTGGGAGGGAGCTAGTGGTGT GGCATGTGTATCCCGCTGGCCAAGCACATGTGAACGAAGCCAAGAATCCA GGGGCTTTTCTGCCAGCCAGCACTGACTCACTTGCGAGGGGCCCTGCCTG GCTCGGGGGAGGCAGGGCTGAAGTACCACATTAGGGCATGTTCCGGGGAA GTAGATTCTCTGAATAACTTGGATGGCTCCCTGGAGCATTTAGGACAGAA GCCACCTGGAAAATAGAGATGGTCACCCCCACGTAGCCTTGACAGTGCCC AGAAAGTCTTGTCACTTGGTAAATGTTAACAGCTATGATCCGTTCTTTAA GACCCTGGGGAGTTTTAAGTTTTACCCCACCAGACCTGAGAAGGGTAAAG GGCTGCAGATTCTGTTCTTTTAACTGGGGCCAGTGTGAGCCATCTTTGAC TCAGTGCTTGCAATAGACCTTGATTCTGCAGTGGGACCTCCCAGGCCCCC TTGCCCCCCGCAACTTCTGGACTCATATCCGTCAGACATCACTTGTCACC TTCCAGCATCAGGGAGAACTGGATCCCTCCTGGCTCCACACTCTTAGGCT CTTTGTAAGTAGCTGGTGAGGGTTTTCTTCTCTCTGCAAGGGAGGCTGGT AGAACTATGGATGTGATTCGTACAATTTTAGAGACAAAAAGAAAGTACCC AGGAGGTCATTTATTTCAGCTGCTTCATTGCATAGGTCGGGGAGTTGAGC ATGGAGTCCAGCAGCTACTAACTAGTTATCTCTGTACCTGGCTTCCATTT ACTGGTCCTTAGCTTGTTCCGTGATTCTTCATTGCCCCTTATTTCTCACC AGAGGGACTGGTTGGCCCTAGATGGAGTGGTCTTTTTAAAATTTTTTTTT TAAATTTTTTGAGACAGAGTCTCACTCTGTCACCTAGGCTGTAGTGCAGT GCTGCGATCTCGGCTCACTGCAACCTCCGCCTCCTGAGTTCAAGCAATTC TCCTGTCTCAGCCTCCTGAGTAGCTGGGATTACAGGTGTGTACCACTATG CCCAGCTAATTTTTGTATTTTTAGTAGAGATGGGATTTCACCATATTGGC CAGGTTGGTCTTGAACTCCTGACCTCAAATGATCTGCCCACCTTAGCCTC CCGAAGTGCTGGGATTGCAGGTGTGAGCCACCGCACCTGGCCTGGGCAGA GTGAAGTCTTATGCTGGGGAGCCATCAGCATGCTCAAACCTCCTGCAATT GTAGCACACTTTGTAAAACTGTTTCCCACAAAAGGGCAGAACTATTTGGG ACTTTCATGAGACCATTCACTTTGTAGCACATACTACTTTGAAGTTTATA CCTTGGAAAACCTCATGATGGTATTCCCAGGCTTGCACGTAATCTGCACT CAAAACATAGCTGTAGAATTGAACTAAAGCATCCCTCTGTCCAATTAAGA CCTATAACCTCTCTTTTTGAGACAGAATCTCGCTCTGTCACCCAGGTTGG AGTGCAGTGGTGCAATCTCAGCTCACTGCATCCTTCGCCTCCTGGATTCA AGCGATTCTCTTGCCTTAGCCTCCGAAGTAACTGGGACTACAGGTGCGCG CCACCACGCCTGGGTAATTTTTGTATTTTTAGTAGAGACGGGGTTTCGCC ATGGCCAGGCTGGTCTCAAACTCCTGGCCTCAAGTGATCCTCCCGCCTCA GCCTCCCAAAGTGCTGGGATTACAGGGTGCACCACCACACCCAGCCAGGA CCTATGATCTAATTCATTGTTGGGGTAGCTTCACAATTTTCTTCTGGACG CCTTAGTAAGTCCACACTTTAAGCAGCCACCACATGGCATACTTTACCTT CTGTTTTTCCTTTCCCCTCCCCTACCTAGACCCTCCTAACTTTTGGGGTT TTTTTCCTTTCCTCAGGGTCAGTTTGAAAAGGAGGATCGAGCTCACTGTG GAGTATCCATGGAGATGTGGAGCCTTGTCACCAACCTCTAACTGCAGAAC TGGG ATG

III. DNA Methylation

In some embodiments, the present invention provides using oligonucleotide that are methylated at specific sites for screening purposes. The present invention is not limited to a particular mechanism. Indeed, an understanding of the mechanism is not necessary to practice the present invention. Nonetheless, it is contemplated that one mechanism for the regulation of gene activity is methylation of cytosine residues in DNA. 5-methylcytosine (5-MeC) is the only naturally occurring modified base detected in DNA (Ehrlick et al., Science 212:1350-1357 (1981)). Although not all genes are regulated by methylation, hypomethylation at specific sites or in specific regions in a number of genes is correlated with active transcription (Doerfler, Annu Rev. Biochem. 52:93-124 [1984]; Christman, Curr. Top. Microbiol. Immunol. 108:49-78 [1988]; Cedar, Cell 34:5503-5513 [1988]). DNA methylation in vitro can prevent efficient transcription of genes in a cell-free system or transient expression of transfected genes. Methylation of C residues in some specific cis-regulatory regions can also block or enhance binding of transcriptional factors or repressors (Doerfler, supra; Christman, supra; Cedar, Cell 34:5503-5513 (1988); Tate et al., Curr. Opin. Genet. Dev. 3:225-231 [1993]; Christman et al., Virus Strategies, eds. Doerfler, W. & Bohm, P. (VCH, Weinheim, N.Y.) pp. 319-333 [1993]).

Disruption of normal patterns of DNA methylation has been linked to the development of cancer (Christman et al., Proc. Natl. Acad. Sci. USA 92:7347-7351 [1995]). The 5-MeC content of DNA from tumors and tumor derived cell lines is generally lower than normal tissues (Jones et al., Adv. Cancer Res 40:1-30 [1983]). Hypomethylation of specific oncogenes such as c-myc, c-Ki-ras and c-Ha-ras has been detected in a variety of human and animal tumors (Nambu et al., Jpn. J. Cancer (Gann) 78:696-704 [1987]; Feinberg et al., Biochem. Biophys. Res. Commun. 111:47-54 [1983]; Cheah et al., JNCI73:1057-1063 [1984]; Bhave et al., Carcinogenesis (Lond) 9:343-348 [1988]. In one of the best studied examples of human tumor progression, it has been shown that hypomethylation of DNA is an early event in development of colon cancer (Goetz et al., Science 228:187-290 [1985]). Interference with methylation in vivo can lead to tumor formation. Feeding of methylation inhibitors such as L-methionine or 5-azacytodine or severe deficiency of 5-adenosine methionine through feeding of a diet depleted of lipotropes has been reported to induce formation of liver tumors in rats (Wainfan et al., Cancer Res. 52:2071s-2077s [1992]). Studies show that extreme lipotrope deficient diets can cause loss of methyl groups at specific sites in genes such as c-myc, ras and c-fos (Dizik et al., Carcinogenesis 12:1307-1312 [1991]). Hypomethylation occurs despite the presence of elevated levels of DNA MTase activity (Wainfan et al., Cancer Res. 49:4094-4097 [1989]). Genes required for sustained active proliferation become inactive as methylated during differentiation and tissue specific genes become hypomethylated and are active. Hypomethylation can then shift the balance between the two states. In some embodiment, the present invention thus takes advantage of this naturally occurring phenomena, to provide compositions and methods for site specific methylation of specific gene promoters, thereby preventing transcription and hence translation of certain genes. In other embodiments, the present invention provides methods and compositions for upregulating the expression of a gene of interest (e.g., a tumor suppressor gene) by altering the gene's methylation patterns.

The present invention describes the use of unmodified completely complementary DNA oligonucleotide sequences to inhibit gene expression. The present invention is not limited to the use of methylated oligonucleotides or modified oligonucleotides to identify therapeutic sequences. We describe the use of non-methylated oligonucleotides for the inhibition of gene expression and we prove this system works by providing the results of experiments conducted during the course of development of the present invention. For example we demonstrate that an unmethylated oligonucleotide targeted toward Bcl-2 inhibited the growth of lymphoma cells to a level that was comparable to that of a methylated oligonucleotide.

IV. Oligonucleotides

The term “oligonucleotide,” refers to a short length of single-stranded polynucleotide chain. Oligonucleotides are typically less than 200 residues long (e.g., between 8 and 100), however, as used herein, the term is also intended to encompass longer polynucleotide chains (e.g., as large as 5000 residues). Oligonucleotides are often referred to by their length. For example a 24 residue or base oligonucleotide is referred to as a “24-mer”. Oligonucleotides can form secondary and tertiary structures by self-hybridizing or by hybridizing to other polynucleotides. Such structures can include, but are not limited to, duplexes, hairpins, cruciforms, bends, and triplexes.

In some embodiments, the present invention provides DNAi oligonucleotides for inhibiting the expression of oncogenes. Exemplary design and production strategies for DNA is are described below. The below description is not intended to limit the scope of DNAi compounds suitable for use in the present invention. One skilled in the relevant recognizes that additional DNA is are within the scope of the present invention.

A. Oligonucleotide Design

In some embodiments, oligonucleotides are designed based on preferred design criteria. Such oligonucleotides can then be tested for efficacy using the methods disclosed herein. For example, in some embodiments, the oligonucleotides are methylated at least one, preferably at least two, and even more preferably, all of the CpG islands. In other embodiments, the oligonucleotides contain no methylation. The present invention is not limited to a particular mechanism. Indeed, an understanding of the mechanism is not necessary to practice the present invention. Nonetheless, it is contemplated that preferred oligonucleotides are those that have at least a 40% CG content and at least 1 CG dinucleotides. In some embodiments, oligonucleotides are designed with at least 1 A or T to minimize self hybridization. In some embodiments, commercially available computer programs are used to survey oligonucleotides for the ability to self hybridize. Preferred oligonucleotides are at least 10, and preferably at least 15 nucleotides and no more than 100 nucleotides in length. Particularly preferred oligonucleotides are 20-34 nucleotides in length. In some embodiments, oligonucleotides comprise the universal protein binding sequences CCGCCC and CGCG or the complements thereof. In some embodiments, oligonucleotides comprise the universal protein binding sequences (G/T)CCCGCCC(G) and the complements thereof. It is also preferred that the oligonucleotide hybridize to a promoter region of a gene upstream from the TATA box of the promoter. It is also preferred that oligonucleotide compounds are not completely homologous to other regions of the human genome. The homology of the oligonucleotide compounds of the present invention to other regions of the genome can be determined using available search tools (e.g., BLAST, available at the Internet site of NCBI).

In some embodiments, oligonucleotides are designed to hybridize to regions of the promoter region of an oncogene known to be bound by proteins (e.g., transcription factors). Exemplary oligonucleotide compounds of the present invention are shown in Table 3. The present invention is not limited to the oligonucleotides described herein. Other suitable oligonucleotides may be identified (e.g., using the criteria described above). Exemplary oligonucleotide variants of the disclosed oligonucleotides can include smaller oligonucleotide sequences of 20-mer or can be right or left shifted 20 base pairs. Candidate oligonucleotides may be tested for efficacy using any suitable method, including, but not limited to, those described in the illustrative examples below. Using the in vitro assay described below in the material and methods and Figures, candidate oligonucleotides can be evaluated for their ability to prevent cell proliferation or target inhibition at a variety of concentrations. Particularly preferred oligonucleotides are those that inhibit gene expression of target proteins as a low concentration (e.g., less that 20 μM, and preferably, less than or equal to 10 μM in the in vitro assays disclosed herein).

B. Materials and Methods

Oligonucleotide Preparation (FIGS. 1-25, 27-30, 31-49, 54-67)

All oligonucleotides were synthesized utilizing cyanoethyl phosphoramidite chemistry, purified by reverse phase high-performance liquid chromatography (RP-HPLC), and lyophilized by The Midland Certified Reagent Company (Midland, Tex.). Methylated oligonucleotides were methylated at all CpG sites.

Cell Culture (FIGS. 1-25, 27-30, 31-49, 54-67)

Human lung carcinoma cells (A549; ATCC) were cultivated in DMEM medium (ATCC) containing 10% fetal bovine serum (FBS; Invitrogen) and maintained under a humidified atmosphere of 5% CO2 at 37° C. Cells were split 1:8 at 90% confluence and used for experiments between passages 12 and 20 (2,500 cells per well were plated 12-24 hours prior to adding oligonucleotides).

Human breast carcinoma cells (MDA-MB-231; ATCC) were cultivated in Leibovitz's L-15 medium (ATCC) containing 10% fetal bovine serum (FBS; Invitrogen) and maintained under a humidified atmosphere at 37° C. Cells were split 1:6 at 90% confluence and used for experiments between passages 15 and 22 (2,500 cells per well were plated 12-24 hours prior to adding oligonucleotides).

Human prostate carcinoma cells (DU145; ATCC) were cultivated in EMEM medium (ATCC) containing 10% fetal bovine serum (FBS; Invitrogen) and maintained under a humidified atmosphere of 5% CO2 at 37° C. Cells are split 1:8 at 90% confluence and used for experiments between passages 10 and 16 (2,500 cells per well were plated 12-24 hours prior to adding oligonucleotides).

Human breast carcinoma cells (MCF-7; ATCC) were cultivated in 50:50 RPMI/DMEM medium (ATCC) containing 10% fetal bovine serum (FBS; Corning), 0.01 mg/mL insulin (Sigma-Aldrich) and maintained under a humidified atmosphere at 37° C. at 5% CO2. Cells were split 1:6 at 90% confluence and used for experiments between passages 15 and 18 (2,500 cells per well were plated 12-24 hours prior to adding oligonucleotides).

Human colorectal carcinoma cells (HCT-116; ATCC) were cultivated in McCoy's 5A medium (Corning) containing 10% fetal bovine serum (FBS; Corning) and maintained under a humidified atmosphere at 37° C. at 5% CO2. Cells were split 1:6 at 90% confluence and used for experiments between passages 4 and 7 (2,500 cells per well were plated 12-24 hours prior to adding oligonucleotides).

HepG2 cells were plated using 5,000 cells per well in 96 well plate (for both qPCR experiment and cell count experiments). Cells were incubated for 24 hours prior to treatment with DNAi oligonucleotides. Twenty-four hours after plating DNAi oligonucleotides were added to the cells at final concentration of 15 uM. At each timepoint (24, 72, and 144 hours) cells from 96 well plate were washed with 1×PBS once and total RNA isolated using MagMax-96 Total RNA isolation kit (Lifetech, cat#AM1830). At 72 hour timepoint cells were over 90% confluent, therefore cells were washed with 1×PBS twice, trypsinized with 0.05% Trypsin-EDTA and transferred from each individual well (96-well plate) into 24-well plate. STAT3 DNAi oligonucleotides were added to the cells in 24-well plate at final concentration of 15 uM.

HepG2 cells were trypsinized (as described above) and cells from each well (96-well plate) were diluted in 1 mL of complete growth medium prior to cell counting performed using Guava PCA-96 flow cytometry system. HepG2 cell culture work was performed at Altogen Labs (Austin, Tex.).

mRNA Expression Analysis and RNA Isolation (FIG. 67)

All RNA was isolated using the MAGMAX96 Total RNA Isolation kit (cat#AM1830; Lifetech). The manufacturer's protocol was followed, including a final elution of 50 μL elution solution. RNA was stored at −20° C. for later use.

Reverse Transcription (RT) (FIG. 67)

Isolated RNA was reverse transcribed into cDNA in a single reaction containing RNase Inhibitor Protein (15518; Lifetech) and MMLV-Reverse Transcriptase (18057; Lifetech). RNA input into the RT reaction was based on a 7.5 μL input per 20 μL reaction size for all samples.

qPCR (FIG. 67)

Fluorescence based, real-time reverse transcription-PCR (qRT-PCR) is a standard tool used for quantification of mRNA levels. This technique has high throughput capabilities with both high sensitivity and specificity for the target of interest. The amplification reaction consisted of dNTPs (PCR grade; Roche) and Platinum Taq Polymerase (10966; Lifetech). Cycling conditions were as follows: 95° C. for 1 minute; then 50 cycles of 95° C. for 5 seconds and 60° C. for 20 seconds. Results were determined by real-time PCR on the ABI Prism 7900 SDS real-time PCR machine (Applied Biosystems, Foster City, Calif.). All qPCR work was performed at Altogen Labs (Austin, Tex.).

As shown in FIG. 67, PC2 (206; exposed at 15 μM), a PCSK9 targeted oligonucleotide, demonstrated an approximate 40% decrease of PCSK9 mRNA at 72 hours post-exposure compared to control PCSK9 mRNA levels in HepG2 cells. While PC2 (206) decreased PCSK9 mRNA expression, it was not cytotoxic to cells at either 24 or 72 hours post-exposure in the same experiment. This demonstrates that an oligonucleotide is capable of modulating target gene expression with expected phenotypic changes.

Altogen Labs (Austin, Tex.) performed the cell culture work for A549, MDA-MB-231, DU145 and START Preclinical (San Antonio, Tex.) performed the cell culture work for MCF-7 and HCT-116.

Cell Growth Inhibition Assay (FIGS. 1-25, 27-30, 31-49, 54-66)

Cells were harvested from T-75 flask by a single wash with 1×PBS and incubation with 2 ml of 0.05% Trypsin-EDTA (Invitrogen) for 7 minutes at 37□C. Trypsin was inactivated by addition of 8 ml of complete medium (total volume of 10 ml). Cells were counted using hemocytometer and cell count confirmed by Guava PCA flow cytometry. Cells were then plated and assayed. Cell growth inhibition was assessed using a Vybrant MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) Cell Proliferation Assay (cat#V13154) purchased from Life Technologies (Carlsbad, Calif.). For each cell line 2,500 cells per well were plated 12 hours prior to adding oligonucleotides. Absorbance measurements at 570 nm were made using a Molecular Devices Spectramax Plus (Sunnyvale, Calif.) microplate reader. Each treatment was run in quadruplicate. Altogen Labs (Austin, Tex.) and START Preclinical (San Antonio, Tex.) performed the cell growth inhibition assay. Included in Tables 4 and 5 are the sequences for the control and negative control oligonucleotides used in the experiments.

Oligonucleotide Preparation (FIGS. 26, 50-53; Descriptions Referenced in U.S. Pat. No. 7,524,827)

All oligonucleotides were synthesized, gel purified anal lyophilized by BIOSYNTHESIS (Lewisville, Tex.) or Qiagen (Valencia, Calif.). Methylated oligonucleotides were methylated at all CpG sites. Methylated Oligonucleotides were dissolved in pure sterile water (Gibco, Invitrogen Corporation) and used to treat cells in culture.

Cell Culture (FIGS. 26, 50-53; Descriptions Referenced in U.S. Pat. No. 7,524,827)

Human breast cancer cells, MCF7 and MDA-MB-231, were obtained from Karmanos Cancer Institute. All cells were cultured in DMEM/F12 media (Gibco, Md.) supplemented with 10 mM HEPES, 29 mM sodium bicarbonate, penicillin (100 units/ml) and streptomycin (100 μg/ml). In addition, 10% calf serum, 10 μg/ml insulin (Sigma Chemical, St Louis, Mo.), and 0.5 nM estradiol was used in MCF7 media and 10% fetal calf serum was used for MDA-MB 231. All flasks and plates were incubated in a humidified atmosphere of 95% air and 5% CO2 at 37° C.

BxPC-3 pancreatic carcinoma cell line was cultured in RPMI 1640 with 10% FBS.

NMuMG (normal mouse mammary gland cells) cell line was grown in DMEM media with 4.5 g/l glucose, 10 μg/ml insulin and 10% FBS.

All the above cells were seeded at 2,500 to 5,000 cells/well in 96 well plates. The cells were treated with oligonucleotide compounds in fresh media (100 μl total volume) 24 hours after seeding. The media was replaced with fresh media without oligonucleotides 24 hours after treatment and every 48 hours for 6 to 7 days or until the control cells were 80 to 100% confluent. The inhibitory effect of oligonucleotide was evaluated using an MTT staining technique.

Cell Growth Inhibition Assay (FIGS. 26, 50-53; Descriptions Referenced in U.S. Pat. No. 7,524,827)

Cell growth inhibition was assessed using 3-[4,5-Dimethyl-thiazol-2-yl]-2,5diphenyltetrazolium bromide (MTT) purchased from Sigma Chemical (St. Louis, Mo.). Cells were resuspended in culture media at 50,000 cells/ml and 100 μl was distributed into each well of a 96-well, flat bottomed plate (Costar Corning, N.Y., USA) and incubated for 24 hours. Media was changed to 100 μl fresh media containing the desired concentration of oligonucleotides and incubated for 24 hours. Controls had media with pure sterile water equal to the volume of oligonucleotide solution. The media was changed without further addition of oligonucleotides every 24 hours until the control cultures were confluent (6 to 7 days). Thereafter the media was removed and plates were washed two times with phosphate-buffered saline (PBS) and 100 μl of serum free media containing 0.5 mg/ml MTT dye was added into each well and incubated for 1 hour at 37° C. The media with dye was removed, washed with PBS and 100 μl of dimethyl sulfoxide (DMSO) was added to solubilize the reactive dye. The absorbance values were read using an automatic multiwell spectrophotometer (Bio-Tek Microplate Autoreader, Winooski, Vt., USA). Each treatment was repeated at least 3 times with 8 independent wells each time. Included in Tables 4 and 5 are the sequences for the control and negative control oligonucleotides used in the experiments.

C. Preparation and Formulation of Oligonucleotides

Any of the known methods of oligonucleotide synthesis can be used to prepare the modified oligonucleotides of the present invention. In some embodiments utilizing methylated oligonucleotides the nucleotide, dC is replaced by 5-methyl-dC where appropriate, as taught by the present invention. The modified or unmodified oligonucleotides of the present invention are most conveniently prepared by using any of the commercially available automated nucleic acid synthesizers. They can also be obtained from commercial sources that synthesize custom oligonucleotides pursuant to customer specifications.

While oligonucleotides are a preferred form of compound, the present invention comprehends other oligomeric oligonucleotide compounds, including but not limited to oligonucleotide mimetics such as are described below. The oligonucleotide compounds in accordance with this invention preferably comprise from about 20 to about 34 nucleobases (i.e., from about 20 to about 34 linked bases), although both longer and shorter sequences may find use with the present invention.

Specific examples of preferred compounds useful with the present invention include oligonucleotides containing modified backbones or non-natural internucleoside linkages. As defined in this specification, oligonucleotides having modified backbones include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone. For the purposes of this specification, modified oligonucleotides that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides.

Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3′-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3′-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′. Various salts, mixed salts and free acid forms are also included.

Preferred modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH2 component parts.

In other preferred oligonucleotide mimetics, both the sugar and the internucleoside linkage (i.e., the backbone) of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar-backbone of an oligonucleotide is replaced with an amide containing backbone, in particular an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is herein incorporated by reference. Further teaching of PNA compounds can be found in Nielsen et al., Science 254:1497 (1991).

In some embodiments, oligonucleotides of the invention are oligonucleotides with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular —CH2, —NH—O—CH2-, —CH2-N(CH3)-O—CH2- [known as a methylene (methylimino) or MMI backbone], —CH2-O—N(CH3)-CH2-, —CH2-N(CH3)-N(CH3)-CH2-, and —O—N(CH3)-CH2-CH2- [wherein the native phosphodiester backbone is represented as —O—P—O—CH2-] of the above referenced U.S. Pat. No. 5,489,677, and the amide backbones of the above referenced U.S. Pat. No. 5,602,240. Also preferred are oligonucleotides having morpholino backbone structures of the above-referenced U.S. Pat. No. 5,034,506.

Modified oligonucleotides may also contain one or more substituted sugar moieties. Preferred oligonucleotides comprise one of the following at the 2′ position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl. Particularly preferred are O[(CH2)nO]mCH3, O(CH2)nOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON[(CH2)nCH3)]2, where n and m are from 1 to about 10. Other preferred oligonucleotides comprise one of the following at the 2′ position: C1 to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties. A preferred modification includes 2′-methoxyethoxy (2′-O—CH2CH2OCH3, also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al., Helv. Chim. Acta 78:486 [1995]) i.e., an alkoxyalkoxy group. A further preferred modification includes 2′-dimethylaminooxyethoxy (i.e., a O(CH2)2ON(CH3)2 group), also known as 2′-DMAOE, and 2′-dimethylaminoethoxyethoxy (also known in the art as 2′-O-dimethylaminoethoxyethyl or 2′-DMAEOE), i.e., 2′-O—CH2-O—CH2-N(CH2)2.

Other preferred modifications include 2′-methoxy(2′-O—CH3), 2′-aminopropoxy(2′-OCH2CH2CH2NH2) and 2′-fluoro (2′-F). Similar modifications may also be made at other positions on the oligonucleotide, particularly the 3′ position of the sugar on the 3′ terminal nucleotide or in 2′-5′ linked oligonucleotides and the 5′ position of 5′ terminal nucleotide. Oligonucleotides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.

Oligonucleotides may also include nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine. Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds of the invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. and are presently preferred base substitutions, even more particularly when combined with 2′-O-methoxyethyl sugar modifications.

Another modification of the oligonucleotides of the present invention involves chemically linking to the oligonucleotide one or more moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety, cholic acid, a thioether, (e.g., hexyl-S-tritylthiol), a thiocholesterol, an aliphatic chain, (e.g., dodecandiol or undecyl residues), a phospholipid, (e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate), a polyamine or a polyethylene glycol chain or adamantane acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety.

One skilled in the relevant art knows well how to generate oligonucleotides containing the above-described modifications. The present invention is not limited to the antisense oligonucleotides described above. Any suitable modification or substitution may be utilized.

It is not necessary for all positions in a given compound to be uniformly modified, and in fact more than one of the aforementioned modifications may be incorporated in a single compound or even at a single nucleoside within an oligonucleotide. The present invention also includes pharmaceutical compositions and formulations that include the antisense compounds of the present invention as described below.

D. Cocktails

In some embodiments, the present invention provides cocktails comprising two or more oligonucleotides directed towards promoter regions of genes (e.g., oncogenes). In some embodiments, the two oligonucleotides hybridize to different regions of the promoter of the same gene. In other embodiments, the two or more oligonucleotides hybridize to promoters of two different genes. The present invention is not limited to a particular mechanism. Indeed, an understanding of the mechanism is not necessary to practice the present invention. Nonetheless, it is contemplated that the combination of two or more compounds of the present invention provides an inhibition of cancer cell growth that is greater than the additive inhibition of each of the compounds administered separately.

V. Research Uses

The present invention is not limited to therapeutic applications. For example, in some embodiments, the present invention provides compositions and methods for the use of oligonucleotides as a research tool.

A. Kits

For example, in some embodiments, the present invention provides kits comprising oligonucleotides specific for inhibition of a gene of interest, and optionally cell lines (e.g., cancer cells lines) known to express the gene. Such kits find use, for example, in the identification of metabolic pathways or the involvement of genes in disease (e.g., cancer), as well as in diagnostic applications. In some embodiments, the kits further comprise buffer and other necessary reagents, as well as instructions for using the kits.

B. Target Validation

In some embodiments, the present invention provides methods and compositions for use in the validation of gene targets (e.g., genes suspected of being involved in disease). For example, in some embodiments, the expression of genes identified in broad screening applications (e.g., gene expression arrays) as being involved in disease is downregulated using the methods and compositions of the present invention. The methods and compositions of the present invention are suitable for use in vitro and in vivo (e.g., in a non-human animal) for the purpose of target validation. In other embodiments, the compounds of the present invention find use in transplantation research (e.g., HLA inhibition).

C. Drug Screening

In other embodiments, the methods and compositions of the present invention are used in drug screening applications. For example, in some embodiments, oligonucleotides of the present invention are administered to a cell (e.g., in culture or in a non-human animal) in order to inhibit the expression of a gene of interest. In some embodiments, the inhibition of the gene of interest mimics a physiological or disease condition. In other embodiments, an oncogene or disease causing gene is inhibited. Test compounds (e.g., small molecule drugs or oligonucleotide mimetics) are then administered to the test cell and the effect of the test compounds is assayed.

The test compounds of the present invention can be obtained using any of the numerous approaches in combinatorial library methods known in the art, including biological libraries; peptoid libraries (libraries of molecules having the functionalities of peptides, but with a novel, non-peptide backbone, which are resistant to enzymatic degradation but which nevertheless remain bioactive; see, e.g., Zuckennann et al., J. Med. Chem. 37: 2678-85 [1994]); spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the ‘one-bead one-compound’ library method; and synthetic library methods using affinity chromatography selection. The biological library and peptoid library approaches are preferred for use with peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds (Lam (1997) Anticancer Drug Des. 12:145).

Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt et al., Proc. Natl. Acad. Sci. U.S.A. 90:6909 [1993]; Erb et al., Proc. Nad. Acad. Sci. USA 91:11422 [1994]; Zuckermann et al., J. Med. Chem. 37:2678 [1994]; Cho et al., Science 261:1303 [1993]; Carrell et al., Angew. Chem. Int. Ed. Engl. 33.2059 [1994]; Carell et al., Angew. Chem. Int. Ed. Engl. 33:2061 [1994]; and Gallop et al., J. Med. Chem. 37:1233 [1994].

Libraries of compounds may be presented in solution (e.g., Houghten, Biotechniques 13:412-421 [1992]), or on beads (Lam, Nature 354:82-84 [1991]), chips (Fodor, Nature 364:555-556 [1993]), bacteria or spores (U.S. Pat. No. 5,223,409; herein incorporated by reference), plasmids (Cull et al., Proc. Nad. Acad. Sci. USA 89:18651869 [1992]) or on phage (Scott and Smith, Science 249:386-390 [1990]; Devlin Science 249:404-406 [1990]; Cwirla et al., Proc. NatI. Acad. Sci. 87:6378-6382 [1990]; Felici, J. Mol. Biol. 222:301 [1991]).

VI. Compositions and Delivery

In some embodiments, the oligonucleotide compounds of the present invention are formulated as pharmaceutical compositions for delivery to a subject as a pharmaceutical. The novel antigen compounds of the present invention find use in the treatment of a variety of disease states and conditions in which it is desirable to inhibit the expression of a gene or the growth of a cell. In some preferred embodiments, the compounds are used to treat disease states resulting from uncontrolled cell growth, for example including, but not limited to, cancer. The present invention is not limited to the treatment of a particular cancer. The oligonucleotide compounds of the present invention are suitable for the treatment of a variety of cancers including, but not limited to, breast, colon, lung, stomach, pancreatic, bladder, leukemia, and lymphoma. In other preferred embodiments, the compounds are used to treat disease states resulting from gene expression, for example including, but not limited to, non cancer diseases. The below discussion provides exemplary, non-limiting examples of formulations and dosages.

A. Pharmaceutical Compositions

The present invention further provides pharmaceutical compositions (e.g., comprising the oligonucleotide compounds described above). The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic and to mucous membranes including vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer); intratracheal, intranasal, epidermal and transdermal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration.

Pharmaceutical compositions and formulations for topical administration may include transdermal patches, needless injectors, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

Compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable.

Compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions that may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, nanoparticle, nanocrystal, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids.

The pharmaceutical formulations of the present invention, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

The compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances that increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension may also contain stabilizers.

In one embodiment of the present invention the pharmaceutical compositions may be formulated and used as foams. Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, jellies and liposomes. While basically similar in nature these formulations vary in the components and the consistency of the final product.

Agents that enhance uptake of oligonucleotides at the cellular level may also be added to the pharmaceutical and other compositions of the present invention. For example, cationic lipids, such as lipofectin (U.S. Pat. No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (WO 97/30731), cochleates (Patent application numbers 20080242625 and 20120294901) also enhance the cellular uptake of oligonucleotides.

The compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the nucleic acid(s) of the formulation.

Compositions and formulations for oral administration include powders or granules, microparticulates, nanoparticulates, suspensions or solutions in water or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable. Preferred oral formulations are those in which oligonucleotides of the invention are administered in conjunction with one or more penetration enhancers surfactants and chelators. Preferred surfactants include fatty acids and/or esters or salts thereof, bile acids and/or salts thereof. Preferred bile acids/salts include chenodeoxycholic acid (CDCA) and ursodeoxychenodeoxycholic acid (UDCA), cholic acid, dehydrocholic acid, deoxycholic acid, glucholic acid, glycholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, sodium tauro-24,25-dihydro-fusidate, sodium glycodihydrofusidate. Preferred fatty acids include arachidonic acid, undecanoic acid, oleic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a monoglyceride, a diglyceride or a pharmaceutically acceptable salt thereof (e.g. sodium). Also preferred are combinations of penetration enhancers, for example, fatty acids/salts in combination with bile acids/salts. A particularly preferred combination is the sodium salt of lauric acid, capric acid and UDCA. Further penetration enhancers include polyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether. Oligonucleotides of the invention may be delivered orally in granular form including sprayed dried particles, or complexed to form micro or nanoparticles or nanocrystals. Oligonucleotide complexing agents include poly-amino acids; polyimines; polyacrylates; polyalkylacrylates, polyoxethanes, polyalkylcyanoacrylates; cationized gelatins, albumins, starches, acrylates, polyethyleneglycols (PEG) and starches; polyalkylcyanoacrylates; DEAE-derivatized polyimines, pollulans, celluloses and starches. Particularly preferred complexing agents include chitosan, N-trimethylchitosan, poly-L-lysine, polyhistidine, polyornithine, polyspermines, protamine, polyvinylpyridine, polythiodiethylamino-methylethylene P(TDAE), polyaminostyrene (e.g. p-amino), poly(methylcyanoacrylate), poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(isohexylcynaoacrylate), DEAE-methacrylate, DEAE-hexylacrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran, polymethylacrylate, polyhexylacrylate, poly(D,L-lactic acid), poly(DL-lactic-co-glycolic acid (PLGA), alginate, phosphatidylserine, calcium, and polyethyleneglycol (PEG).

Certain embodiments of the invention provide pharmaceutical compositions containing (a) one or more oligonucleotide compounds and (b) one or more other chemotherapeutic agents that function by a non-oligonucleotide mechanism. Examples of such chemotherapeutic agents include, but are not limited to, cytotoxic agents, small molecule protein inhibitors, antibodies, and anti-sense anticancer drugs such as daunorubicin, dactinomycin, doxorubicin, bleomycin, mitomycin, nitrogen mustard, chlorambucil, melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine (CA), 5-fluorouracil (5-FU), floxuridine (5-FUdR), methotrexate (MTX), colchicine, vincristine, vinblastine, etoposide, teniposide, cisplatin, lenalomide, and diethylstilbestrol (DES). Anti-inflammatory drugs, including but not limited to nonsteroidal anti-inflammatory drugs and corticosteroids, and antiviral drugs, including but not limited to ribivirin, vidarabine, acyclovir and ganciclovir, may also be combined in compositions of the invention. Other non-oligonucleotide chemotherapeutic agents are also within the scope of this invention. Two or more combined compounds may be used together or sequentially.

B. Delivery

The oligonucleotide compounds of the present invention may be delivered using any suitable method. In some embodiments, naked DNA is administered. In other embodiments, lipofection is utilized for the delivery of nucleic acids to a subject. In still further embodiments, oligonucleotides are modified with phosphothioates for delivery (See e.g., U.S. Pat. No. 6,169,177, herein incorporated by reference).

In some embodiments, nucleic acids for delivery are compacted to aid in their uptake (See e.g., U.S. Pat. Nos. 6,008,366, 6,383,811 herein incorporated by reference). In some embodiment, compacted nucleic acids are targeted to a particular cell type (e.g., cancer cell) via a target cell binding moiety (See e.g., U.S. Pat. Nos. 5,844,107, 6,077,835, each of which is herein incorporated by reference).

In some embodiments, oligonucleotides are conjugated to other compounds to aid in their delivery. For example, in some embodiments, nucleic acids are conjugated to polyethylene glycol to aid in delivery (See e.g., U.S. Pat. Nos. 6,177,274, 6,287,591, 6,447,752, 6,447,753, and 6,440,743, each of which is herein incorporated by reference). In yet other embodiments, oligonucleotides are conjugated to protected graft copolymers, which are chargeable” drug nano-carriers (PharmaIn). In still further embodiments, the transport of oligonucleotides into cells is facilitated by conjugation to vitamins (Endocyte, Inc, West Lafayette, Ind.; See e.g., U.S. Pat. Nos. 5,108,921, 5,416,016, 5,635,382, 6,291,673 and WO 02/085908; each of which is herein incorporated by reference). In other embodiments, oligonucleotides are conjugated to nanoparticles (e.g., NanoMed Pharmaceuticals; Kalamazoo, Mich.).

In preferred embodiments, oligonucleotides are enclosed in lipids (e.g., liposomes or micelles) to aid in delivery (See e.g., U.S. Pat. Nos. 6,458,382, 6,429,200; each of which is herein incorporated by reference). Preferred liposomes include, but are not limited to amphoteric liposomes (e.g., SMARTICLES,). In still further embodiments, oligonucleotides are complexed with additional polymers to aid in delivery (See e.g., U.S. Pat. Nos. 6,379,966, 6,339,067, 5,744,335; each of which is herein incorporated by reference and Intradigm Corp., Rockville, Md.). Cochleates see e.g. Patent application number: 20080242625 and 20120294901.

In still further embodiments, the controlled high pressure delivery system developed by Mirus (Madison, Wis.) is utilized for delivery of oligonucleotides.

C. Dosages

Dosing is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. In some embodiments, the oligonucleotide is introduced to the host animal at a dosage of between 0.1 mg to 10 g, and preferably at a dosage of between 00.1 mg to 100 mg per kg of body weight or 1 to 300 mg per meter squared body surface area. The administering physician can determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual oligonucleotides, and the delivery means, and can generally be estimated based on EC50s found to be effective in in vitro and in vivo animal models or based on the examples described herein. In general, dosage is from 10 mg to 10 g per kg of body weight, and may be given once or more daily, weekly, monthly or yearly. In some embodiments, dosage is continuous (e.g., intravenously) for a period of from several minutes to several days or weeks. In some embodiments, treatment is given for a defined period followed by a treatment free period. In some embodiments, the pattern of continuous dosing followed by a treatment free period is repeated several times (e.g., until the disease state is diminished).

The treating physician can estimate repetition rates for dosing based on measured residence times and concentrations of the drug in bodily fluids or tissues. Following successful treatment, it may be desirable to have the subject undergo maintenance therapy to prevent the recurrence of the disease state, wherein the oligonucleotide is administered in maintenance doses, ranging from 10 mg to 10 g, preferably from 1 mg to 5 mg, and even more preferably from 0.1 mg to 30 mg per kg of body weight or 0.1 mg/m² to 200 mg/m², once or more daily, to once every 20 years.

VII. Customized Patient Care

In some embodiments, the present invention provides customized patient care.

The compositions of the present invention are targeted to specific genes unique to a patient's disease (e.g., cancer). For example, in some embodiments, a sample of the patient's cancer or other affected tissue (e.g., a biopsy) is first obtained. The biopsy is analyzed for the presence of expression of a particular gene (e.g., oncogene). In some preferred embodiments, the level of expression of an gene in a patient is analyzed. Expression may be detected by monitoring for the presence of RNA or DNA corresponding to a particular oncogene. Any suitable detection method may be utilized, including, but not limited to, those disclosed below. 5 10 15 20

Following the characterization of the gene expression pattern of a patient's gene of interest, a customized therapy is generated for each patient. In preferred embodiments, oligonucleotide compounds specific for genes that are aberrantly expressed in the patient (e.g., in a tumor) are combined in a treatment cocktail. In some embodiments, the treatment cocktail further includes additional chemotherapeutic agents (e.g., those described above). The cocktail is then administered to the patient as described above.

In some embodiments, the analysis of cancer samples and the selection of oligonucleotides for a treatment compound is automated. For example, in some embodiments, a software program that analyses the expression levels of a series of oncogenes to arrive at the optimum selection and concentration of oligonucleotides is utilized. In some embodiments, the analysis is performed by the clinical laboratory analyzing the patient sample and is transmitted to a second provider for formulation of the treatment cocktail. In some embodiments, the information is transmitted over the Internet, thus allowing for the shortest possible time in between diagnosis and the beginning of treatment.

A. Detection of RNA

In some embodiments, detection of oncogenes (e.g., including but not limited to, those disclosed herein) is detected by measuring the expression of corresponding mRNA in a tissue sample (e.g., cancer tissue or other biopsy). In other embodiments, expression of mRNA is measured in bodily fluids, including, but not limited to, blood, plasma, lymph, serum, mucus, and urine. In some preferred embodiments, the level of mRNA expression in measured quantitatively. RNA expression may be measured by any suitable method, including but not limited to, those disclosed below.

In some embodiments, RNA is detected by Northern blot analysis. Northern blot analysis involves the separation of RNA and hybridization of a complementary labeled probe. In other embodiments, RNA expression is detected by enzymatic cleavage of specific structures (INVADER assay, Third Wave Technologies; See e.g., U.S. Pat. Nos. 5,846,717, 6,090,543; 6,001,567; 5,985,557; and 5,994,069; each of which is herein incorporated by reference). The INVADER assay detects specific nucleic acid (e.g., RNA) sequences by using structure-specific enzymes to cleave a complex formed by the hybridization of overlapping oligonucleotide probes.

In still further embodiments, RNA (or corresponding cDNA) is detected by hybridization to a oligonucleotide probe). A variety of hybridization assays using a variety of technologies for hybridization and detection are available. For example, in some embodiments, TaqMan assay (PE Biosystems, Foster City, Calif.; See e.g., U.S. Pat. Nos. 5,962,233 and 5,538,848, each of which is herein incorporated by reference) is utilized. The assay is performed during a PCR reaction. The TaqMan assay exploits the 5′-3′ exonuclease activity of the AMPLITAQ GOLD DNA polymerase. A probe consisting of an oligonucleotide with a 5′-reporter dye (e.g., a fluorescent dye) and a 3′-quencher dye is included in the PCR reaction. During PCR, if the probe is bound to its target, the 5′-3′ nucleolytic activity of the AMPLITAQ GOLD polymerase cleaves the probe between the reporter and the quencher dye. The separation of the reporter dye from the quencher dye results in an increase of fluorescence. The signal accumulates with each cycle of PCR and can be monitored with a fluorimeter.

In yet other embodiments, reverse-transcriptase PCR (RT-PCR) is used to detect the expression of RNA. In RT-PCR, RNA is enzymatically converted to complementary DNA or “cDNA” using a reverse transcriptase enzyme. The cDNA is then used as a template for a PCR reaction. PCR products can be detected by any suitable method, including but not limited to, gel electrophoresis and staining with a DNA specific stain or hybridization to a labeled probe. In some embodiments, the quantitative reverse transcriptase PCR with standardized mixtures of competitive templates method described in U.S. Pat. Nos. 5,639,606, 5,643,765, and 5,876,978 (each of which is herein incorporated by reference) is utilized.

In yet other embodiments, mRNA or transcript numbers are measured using branched DNA technology (e.g. QuantiGene). Branched DNA (bDNA) quantitatively measures gene expression by a sandwich nucleic acid hybridization method that uses bDNA probes specific to the target RNA. The signal from captured target RNA is amplified and enhances assay sensitivity thereby eliminating the need to amplify target RNA by traditional PCR-based gene expression techniques. Furthermore, bDNA assays measure RNA directly from the sample source, without RNA purification or enzymatic manipulation, potentially avoiding inefficiencies and variability introduced by errors inherent to these processes.

B. Detection of Protein

In other embodiments, gene expression of oncogenes is detected by measuring the expression of the corresponding protein or polypeptide. In some embodiments, protein expression is detected in a tissue sample. In other embodiments, protein expression is detected in bodily fluids. In some embodiments, the level of protein expression is quantitated. Protein expression may be detected by any suitable method. In some embodiments, proteins are detected by their binding to an antibody raised against the protein. The generation of antibodies is well known to those skilled in the art.

Antibody binding is detected by techniques known in the art (e.g., radioimmunoassay, ELISA (enzyme-linked immunosorbant assay), “sandwich” immunoassays, immunoradiometric assays, gel diffusion precipitation reactions, immunodiffusion assays, in situ immunoassays (e.g., using colloidal gold, enzyme or radioisotope labels, for example), Western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays, etc.), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc.

In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many methods are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

In some embodiments, an automated detection assay is utilized. Methods for the automation of immunoassays include those described in U.S. Pat. Nos. 5,885,530, 4,981,785, 6,159,750, and 5,358,691, each of which is herein incorporated by reference. In some embodiments, the analysis and presentation of results is also automated. For example, in some embodiments, software that generates an expression profile based on the presence or absence of a series of proteins corresponding to oncogenes is utilized.

In other embodiments, the immunoassay described in U.S. Pat. Nos. 5,599,677 and 5,672,480; each of which is herein incorporated by reference.

VIII Listing of DNAi Sequences

The following sequences in Table 3 are provided as additional non-limiting examples of preferred embodiments of the invention.

TABLE 3 New DNAi Sequences Location relative to 5′ upstream region from Design Sequence gene start Target ID ID No: Sequence (5′-3′) site Survivin SU1 1 GAGCGCACGCCCTCTTAGGCGG 73 Survivin SU2 75 CACCCCGAGGTACGATCAGTGCGTACC 2990 Survivin SU1_02 155 GAGCGCACGCCCTCTTAGGCG 73 Survivin SU1_03 229 GAGCGCACGCCCTCTTAGGCGGTCCA 73 Survivin 303 GTCGCCCCTGGGTCCTGCTGATTGGC 1918 Survivin 322 CAGCGAGCCTGGGCCCCATCGGCACATCT 2905 Survivin 357 CCCGCGGCCTTCTGGGAGTAGAGGC 102 Survivin 431 TCCCGGCGAGTACATCGTTGACTGCACG 675 Survivin 481 AACCTCCTCCCCGCCACGGGTT 1229 Beclin-1 BE1 515 CGACGCCCTTGACCTCCGGCCCGGGGT 39 Beclin-1 BE2 550 CTGCGCCGTTCCCTCTAGGAATGG 111 Beclin-1 572 GAAGCGACGCCCTTGACCTCCGGCCCGG 35 Beclin-1 607 CCCCCGATGCTCTTCACCTCGGG 261 Beclin-1 712 CGGGTCGGCCCCGGAGCGAGGCC 335 Beclin-1 817 GCCCGGCAGCGGCCCCCAGAGGCCG 475 Beclin-1 847 CGGTCTACCGCGGAGGCACTGTGGCCTCGG 308 Beclin-1 952 ACAAAAACTAGCCGGGCGTGGTGGGGCACGCC 735 STAT3 ST1 984 GGCCGAGGCACGCCGTCATGCA −18 STAT3 ST2 985 CCGGCCCTTGGCACCACGTGGTGGCGA 345 STAT3 986 TTGTTCCCTCGGCTGCGACGTCG −135 STAT3 987 CAGTCTGCGCCGCCGCAGCTCCGG −92 STAT3 988 CAGTGCGTGTGCGGTACAGCCG 45 STAT3 989 TGTGCTGGCTGTTCCGACAGTTCGGT 140 STAT3 990 TAACTACGCTATCCCGTGCGGCC 1998449 STAT3 991 TCGCCCAGCCCCAGCCTGGCCGAGGC −35 HIF1A HI1 992 CAGGCCGGCGCGCGCTCCCGCAA 390 HIF1A HI2 1048 GGACGGGCTGCGACGCTCACGTGC 539 HIF1A 1090 GAGGTGGGGGTGCGAGGCGGGAAACCCCTCG 108 HIF1A 1129 CAATCGCCGGGGTCCGGGCCCGGC 162 HIF1A 1130 TGGCCGAAGCGACGAAGAGGG 232 HIF1A 1142 GGGCGGAGGCGCGCTCGGGCGCG 325 HIF1A 1214 CACGGCGGGCGGCCCCCAGGCTCGC 26 HIF1A 1270 CAGGCCGGCGCGCGCTCCCGCAAGCCCG 390 HIF1A 13680 CGATTGCCGCCCAACTCTGCTGGG 789 IL-8 IL8-1 1314 ACGTCCCATTCGGCTCCTGAGCCA 2868 IL-8 IL8-3 1331 GACGTTGACGAAGTCTATCACCCAA 2939 IL-8 1341 ACGGAGTATGACGAAAGTTTTC 257 IL-8 1342 GAGCGAGACTCCCGTCTAAA 3259 KRAS 1535 GCCGGGCCGGCTGGAGAGCGGGTC 5803 KRAS 1538 TCGCCCCTCCTCCGAGACTTTC 6626 KRAS 1584 GCACCCCGCCACCCTCAGGGTCGGC 6029 KRAS 1633 GAGCCGCCGCCACCTTCGCCGCCGC 5475 KRAS 1697 CGGCATAGTTCCCCGCCTTAC 2002 KRAS KR16 1730 CGGCCCGAGCCTCCGTGACGAGTGC 146348 KRAS KR17 1767 CTGGGAGGGGATCCCTCACCGAGAG 3328 MTTP 1784 AACCGCCGTAGCCTCCACTGCG 28 MTTP 1870 TGGCCGCAGTTCGATGACGTAAGACG 1 ApoC-III 1956 GAGTCGGTGGTCCAGGAGGGGCCGC 939 ApoC-III 1957 CTGCGGCTGAGGTGTCATTCGTGACTCAG 3539 ApoC-III 1992 GCGGGCGGGTGAGACAGAAGCGCC 3455 ApoC-III 1993 CCTCGCGAGCGTGGGTGCACGC 3310 ApoC-III 2028 CGATGTCTCCCTCGAGATCACA 3042 ApoC-III 2054 GGACGGACGGATATCTGAGGCCAG 1520 ApoC-III 2062 CGTCCCCGCCACGTTGAAAGGC 3279 ApoC-III 2089 TCTCGGACATGCTCAAATGGTGCAGGCG 3405 ApoC-III 2108 CACCGACAGGAGCCAATAGTGCAACG 4201 ApoC-III 2127 GTCCGGCAGAGGGACCCATGCTGACG 4265 ApoC-III 2136 CGTGAGGCACATGTCCGTGTG 2836 ApoC-III 2170 CAGATGCAGCAAGCGGGCGGGAGAG 123 ApoC-III 2176 CCACGCTGCTGTCCCGCCAGCCCTGCAG 173 ApoC-III 2206 ACCCGCCCCCACCCTGTGTGCCCCC 601 ApoC-III 2225 CGCTCAGAGCCCGAGGCCTTTG 677 ApoB 2252 CGGTGGGGCGGCTCCTGGGCTGC 10 ApoB 2329 CCTCGCGGCCCTGGCTGGCTGGGCG 46 ApoB 2406 AACCGAGAAGGGCACTCAGCCCCG 88 ApoB 2440 CGGCGCCCGCACCCCATTTATAGG 136 ApoB 2451 GTCCAAAGGGCGCCTCCCGGGCC 195 ApoB 2475 CGTCTTCAGTGCTCTGGCGCGGCC 341 ApoB 2513 CACCGGAAGCTTCAGCCAGCGCTCGCTG 988 ApoB 2552 CGAGTGGGAGGCGGCCAGGAGCAAGCCG 1281 ApoB 2553 CGTACACTCACGGAAATGCTGTAAAG 2533 ApoB 2576 CGTCACAGCCAATAATGAGCGTACGC 4862 IL17 2601 CTTGTTTGTATCCGCATGGCTGTGCTC 4451 IL17 2616 CGAGACCGTTGAGGTGGAGTG 3148 IL17 2635 GGTCACTTACGTGGCGTGTCGC 107 IL17 2664 GACAAAATGTAGCGCTATCG 55 MMP2 2666 GCTCCCTGGCCCCGCGCGTCGC 9 MMP2 2732 CCGCGGCGCAGGGCTGCGCTCCGAG 85 MMP2 2865 GCCGCCTGCTACTCCTGGCCTC 453 MMP2 2869 GCGCACTCGGGCCCGCCCCTCTCTGCCC 361 MMP2 2891 CGCTCCGAGGGTCCGCTGGCTCGG 101 MMP2 3024 GTCCACCCTCAGTGCACGACCTCGT 478 MMP2 3066 CACCGCCTGAGGAAGTCTGGATGC 239 MMP2 3101 TGCCTCTCTCGCGATCTGGGCG 512 MMP2 3131 GAGGGACGCCGGCTTGGCTAGGAC 618 FAP 3154 CAGAGCGTGGGTCACTGGATCT 39 FAP 3171 CACCAACATCTGCTTACGTTGAC 272 FAP 3177 TCCACGGACTTTTGAATACCGTGC 133 P-selectin 3184 TAGCTACGAATAAAGAAATTTGTAG 2694 IL6 3185 CACCGCGTGGCTTCTGCCACTTTC 723 IL6 3206 TACGGACGCAGGCACGGCTCTAG 1117 IL6 3226 CAGCTCCGCAGCCGTGCACTGTG 1722 IL6 3255 CTTCACCGATTGTCTAAACAGAGAC 1525 IL6 IL6_1 3256 TTCGTTCCCGGTGGGCTCGAGGGC 35 IL6 3276 TGCTTCCGCGTCGGCACCCAAG 1150 IL23 3300 TCCCTGCATTGTAAGGCCCGCC 195 IL23 3319 CACAGCGGGGATGGGGTGGGAGGG 414 IL23 3320 GACGTCAGAATGAGGCCATCG 1296 IL23 3341 GAGCCAGCACGGTGGTGGGCGCC 1651 IL23 3365 GCGTTTGTCCCACCGGCGCCCCG 4861 IL23 3479 TAACGCCACCCAACAAGTCCGGCG 4830 AKT1 3593 GAGGCTCCCGCGACGCTCACGCG 8 AKT1 3646 TACCGGGCGTCTCAGGTTTTGCC 843 AKT1 3669 TCCGAGCCGCGCACGCCTCAGGC 1562 AKT1 3703 CACCAACGGACTCCGTCCGCCC 2010 AKT1 3770 CCGCCGGCTGCCTCGCTGGCCCAGCG 2464 AKT1 3927 TCTCGGGTCCCGGCCTCGCCCGGCGGAGC 2556 AKT1 4084 CATTCTGGCGGCGCCGCGGCTCGCG 2730 AKT1 4228 CACCGGGCCGCCGCGTCCGGGCGCG 2838 AKT1 AKT4 4338 CACATCCGCCTCCGCCGCCCGG 3160 CRAF 4339 GCGCGAGCCCTACTGGCAGTCG 390 CRAF 4462 CGGGGCGTGGCCTAGCGATCTGGTGGCCG 467 CRAF 4517 TTTCGAAGCTGAAGAGGTTAGGCGACG 499 CRAF 4519 CGACGCTGACTTGCTTTCAGGAG 521 CRAF 4533 AATCGAGAAGAACCGGCTTTCGG 555 CRAF 4556 CTTTGACGCGTCCTCTCCGGGC 689 CRAF 4585 CGGCTCCGCCACTTGACAGCTATGTGG 728 CRAF 4605 AGGCGGAGATTGCGGTGAGCCGAAATCGCG 1582 CRAF 4609 AGGCCGCCCCAACGTCCTGTCGTTCGGCGG 12 CRAF 4677 TCTCGCCCGCTCCTCCTCCCCGCGGCGGGTG 47 CRAF 4745 CGGGAGGCGGTCACATTCGGCGCG 84 CRAF 4782 CGGAGCCCCGAGCAGCCCCCGCATCG 124 CRAF 4871 CGCGCTCCGCGCCTCAGGGCACGCGCC 157 CRAF 4960 AGCCGTTCCCGCCTCACAATCG 234 CRAF 4984 CCGCCATCTAAGATGGCGGCC 270 CRAF 5047 CGGGCGGCCCAGACGAGCGAGCCCTCG 314 CRAF 5110 CGTCCTCCCGACCTGCGACGCCACCGGC 351 Beta- 5233 CGCATATTACTGGGTAAACTCTGTG 1411 catenin Beta- 5234 CACGCTGGATTTTCAAAACAGTTG 5 catenin PCSK9 5235 CAGGGCGCGTGAAGGGGCGCGCGG 120 PCSK9 5236 GACGCGTCCCGGCCCGCCCGAGC 179 PCSK9 5285 GACGCCTGGGGCGCGCAGATCAC 341 PCSK9 5341 CAGGCCGGCGCCCTAGGGGCTCC 494 PCSK9 5359 CACGCCGGCGGCGCCTTGAGCC 56 PCSK9 5402 CAGGTTTCGGCCTCGCCCTCCC 408 PCSK9 5445 CATCGAGCCCGCCATCGCAGCAC 1307 PCSK9 5473 GAGCGCCTCGACGTCGCTGCGGAAACC 273 MEK1 5534 CAAGTCCGGGCCGCGGGCCCCGGGGC 93 MEK1 MEK1_2 5716 GCGCCCCGCGCGGTCCCGTCAGCGC 133 MEK1 5898 GCGGAGCGGGCTGAACGTGCG 249 MEK1 5900 GACTGGAGGCCGGGGGAGGGGCGGGG 433 MEK1 5901 GACCCGGGTAACGCGCTTCCAAC 5 MEK1 MEK1_1 5924 CACTCGGCTCCGCCCCTATTGC 507 MEK1 6000 TACGTCACGGGAGCGCGGCGCAC 578 MEK1 6077 GTCGCGGACGCCGTGGCGCCCTCTGTC 619 MEK1 6154 CACTCGCCGTCATGCCCGGATCC 1183 MEK2 6182 CGCCGCAGCCCGAGTCCGAGAGG 226 MEK2 6202 GAGGGGCGCTGGGGCTGAGGCGAGCG 165 MEK2 6203 CTCGCGATAACGGGATCGGGAGCCGCG 290 MEK2 MEK2_1 6235 CCGACGCGAGGCGGTGCCGGGACCGG 391 MEK2 6240 CACGGCGCGTGTGCCCAAGCGC 436 MEK2 6299 CGTGGACACACGCCCCTAGCCC 643 MEK2 6341 TAGACACTTCGGTGAATCGTGCCGC 1622 CD4 6373 GAGCCACTGCGCCCGGCCTCATTAAGGGCAT 1818 CD4 6406 CGAACAACTTCATTACAATTCGACAAGCGC 2632 CD4 6407 CGTAGTTAAGCGTGTACCAGCCCAAGGC 2522 CD4 6421 GAGCGGTGACCGTGTCTGTCTTAG 3084 CD4 6447 CGGTTTGCAGATTCCAGACCCGATGGACG 4433 WNT1 6466 CGCGCGCCCGCCTCACTCAGCTGAGCG 442 WNT1 6537 CGTCATTCTGTTGCCCTTTGTACCTCG 1226 WNT1 6545 CGCCACGGGCGCATCCATCCCTCCTGGG 4454 WNT1 6579 CACCGCCCTCTAGCCGCCTGCGGG 4960 WNT1 6580 TTGCGGCGACTTTGGTTGTTGCCCGCGACGGT 34 Clusterin 6636 CGTCCCGCCCACCTGCTGCCTGCAGCAG 78 Clusterin 6660 CGACAATCAGCGAGGCACACAGGCT 330 Clusterin 6689 CGGAGAGTAGAGAGGGTTCGCAGTGGCCC 718 Clusterin 6690 CCACGGGGCACAGGCCATAGCCCCG 890 Clusterin 6709 CTCGTGCTCTCAGGCGGCGGTTGCGCCG 3865 Clusterin 6752 CCGGGAGGTGGGGGCCGGTGCAGCACCGG 4260 Clusterin 6753 TCGCGTGCCCATCTGGGAGCCCCTCTCACG 4395 NRAS 6774 CCCCGCCCTCAGCCTAAGCAATGGA 234 NRAS 6793 GACCCCGGAACCGCCATGAACAGCCC 559 NRAS 6818 CCCGCTACGTAATCAGTCGGCGCCCCA 613 NRAS 6961 AACGCAAAAACACCGGATTAATATCGGCCT 142 NRAS 6963 ATAAACGGCCTCTTTACCCAGAGATCA 850 NRAS 6971 CGCCACCTTAAGTTTTTCCAGGCTGC 1779 EZH2 EZH2_2 6986 TCCCGACAAGGGGTGACAGAGGC 1002 EZH2 7002 CGTGAATTCAAGAGTTGCTTAGGCC 1059 EZH2 7003 GACTACCGGTGCCCGCCACCACGCCAGGC 2856 EZH2 7035 GACCGCCCCCCGCCAACCCCACAGCGG 3459 HDAC1 7075 CGCCTCCCGTCCCTACCGTCAGTCGGT 7 HDAC1 7141 CGGTCCGTCCGCCCTCCCGCCCGCGG 30 HDAC1 7207 CGCCAACTTGTGGTCCTACAGTCAACAAG 1740 HDAC1 7226 CGCAGACACGGGCCCGGAACTCGG 173 HDAC1 7258 CGCCCGGCCTAGGAGGGCAGGTTTCTC 1252 PD-1 7297 TGCCGCCTTCTCCACTGCTCAGGCG 23 PD-1 7316 ACCGCCTGACAGCTGGCGCGGCTGCCTGGC 1061 PD-1 PD1 7379 CTGCGAGGCGCGGCCACGGCG 1171 PD-1 7396 CGAGGAGGAAAGGCAGGCGGAGTCCG 3395 PD-1 7397 CAGCGAAGCTGCAGAACGTCCCCATCACCACG 4268 PD-1 7439 CGACAGCCGTGGGAAGGTGCAGTACG 4388 PD-1 7440 CGGGATTCCCTGGAGATGCCTCCAGCGCG 4422 PD-1 7466 AGGCGGTCCCAGGGCTCAGGTGTGGG 2229 PD-1 7498 GCGTGCACCCCGTGGCCAGCTC 3813 PD-1 7526 CAACGTACACGCAATCCACAAC 2832 TNFa 10095 CGGGGAAAGAATCATTCAACCAGCGG 254 TNFa TNF1 10096 CGGTTTCTTCTCCATCGCGGGGGCG 350 TNFa 10129 CTGCTCCGATTCCGAGGGGGGTCTTCT 438 TNFa 10154 CTCCGTGTGGGGCTCTGGTCGGCAGCT 1490 TNFa 10207 CGCAGCCCCGTGGTACATCGAGTGCAGC 2178 MIF1 12470 GACCCGCGCAGAGGCACAGACGC 42 MIF1 12490 CGCCACCGCCGGCGCCAGGCCCCGCCCCCGCG 143 MIF1 12701 CGTTCCTCCAGCAACCGCCGCTAAGCCCGGCG 258 MIF1 12912 CGCCTGCCTCGGCTCGACCCCCGCAG 202 MIF1 13123 CGGCTAGAAATCGGCCTGTTCCGGCCTCGCCT 317 MIF1 13174 CGGGGGTGGGGATGCGGCGGTGAACCCG 404 MIF1 13175 CGCGGCAGGTGAGAGGGGAGCTGCCCCTGCG 588 MIF1 13176 CGCGTGCACGTGTGTCCACATGAGTGC 3676 MIF1 MIF1_1 13203 CGCCACCGCCGGCGCCAGGCCCCGCC 137 MIF1 MIF1_2 13414 CGCGGCAGGTGAGAGGGGAGCTGCCC 583 TTR 11359 CAACGCCCTGGCTCGAGTGCAGTGGCACG 803 TTR 11432 CTACTATCTCAGATACTCGGCCAACTCG 1776 TTR 11450 CACGCGTTTCAGCACTGCACCCTGTTG 2112 HBV 9179 CCGATTGGTGGAGGCAGGAGGAGG 72 HBV 9180 CGAGATTGAGATCTTCTGCGACGCGG 780 HBV 9235 CGCGGCGATTGAGACCTTCGTC 801 HBV 9290 CGTCTGCGAGGCGAGGGAGTTCTTCT 819 HBV 9345 CGATACAGAGCAGAGGCGGTGT 1200 HBV 9346 CGCGTAAAGAGAGGTGCGCCCCGTGG 1674 HBV 9360 ACGGGTCGTCCGCGGGATTCAGCGCCG 1754 HBV 9409 CGTCCCGCGCAGGATCCAGTTGG 1800 HBV 9432 CGGCTGCGAGCAAAACAAGCTGCTAG 1909 HBV 9468 CGCATGCGCCGATGGCCTATGGCCAA 1978 HBV 9496 CGCCGCAGACACATCCAGCGATA 2826 HBV 9525 GCTCCAGACCGGCTGCGA 1900 HBV 9561 CGTCCATCGCAGGATCCAGTTGG 1800 HBV 9562 CGCCGCAGACACATCCAGCGATA 2826 HBV 9591 CAAATGGCACTAGTAAACTGAG 2524 HBV 9592 GAGATTGAGATCTGCGGCGACGCGG 780 HBV 9593 CGACGCGGCGATTGAGATCTTCGTCTG 801 HBV 9594 AGGGGTCGTCCGCGGGATTCAGCGCCG 1754 HAMP 8999 CGTGCCGTCTGTCTGGCTGTCCCAC 1 HAMP 9005 CGAGTGACAGTCGCTTTTATGGGGC 60 HAMP 9035 CGGGGCATGGCCAGCAGCCGCCAGG 424 HAMP 9086 CGTGTGCCCGATCCGCACGTGGTGT 563 HAMP 9121 CGACAGGCTGACGGGCCAAGCTTGG 2344 HAMP 9150 CGGATGGGCAGGGAGGATACCGTTT 3109 HAMP 9151 CGTGGGCGGCGGCGGCTGCGTGGTG 3287 ERBB2 13415 CGGGAAGAGGATGCGCTGACCTGGC 2571 ERBB2 13416 CACGCCCTGGGGAGGAGGCTCGAGAGG 3267 ERBB2 13437 CGAGAGGGGCCGAGCCTCTGAAAAA 3287 ERBB2 13452 CGTCTGGTCCACAGTCCGATGTCCA 3944 PARP1 9595 CCGCCAAAGCTCCGGAAGCCCGACGCC 14 PARP1 9741 CCGCCTCGCCGCCTCGCGTGCGCTC 60 PARP1 9887 CGGGAACGCCCACGGAACCCGCGTC 177 PARP1 9933 CGGGTGGAGCTCTGCGGGCCGCTGC 269 PARP1 9992 CGCCGGCCCCAAACTCTTAAGTGTG 696 PARP1 10014 CGGGAAGCGCAGGCCCCCGCCTCGG 749 PARP1 10045 CGTTCTAACCTGCCGTCCACAGACC 839 ITGA4 10244 GCGCTCTCGGTGGGGAACATTCAACAC 1 ITGA4 10252 CGGGATGCGACGGTTGGCCAACGG 54 ITGA4 10278 CGCAGCGTGTCCGGCGCCAGCGGGC 102 ITGA4 10299 CGGCCCACCGCGGGCGGAGCGTTCG 160 ITGA4 10449 CGCGCACTCGCCCGGCCCCACTCCCG 201 ITGA4 10599 CGCCAGCCGGGAGCTTCGGGTGCTCGCG 235 ITGA4 10749 CGGGTACGGGCCGCTGGGTGGGGTCCCG 272 ITGA4 10899 GTGCGGAGGCGCAGGGCCGGGCTCCG 306 ITGA4 10900 CTACGCGCGGCTGCAGGGGGCGC 339 ITGA4 10938 CTGCGCAGGACTCGCGTCCTGGCCCG 375 ITGA4 11009 CCCGCAGAGCGCGGGATGGCTC 411 ITGA4 11080 CGGACCTGATGGGGCACGGGCTTCCCC 448 ITGA4 11117 CGGTGGTTGGGGCCTAGAAGCG 481 ITGA4 11154 CGCGCCCCTCGCTGTGACCGCCCAGCCCG 524 ITGA4 11203 CGGGGAGTGGGACTGCGGCGGGGAGCCG 580 ITGA4 11208 ACTCGCCGAAGGCCCCTGGGGAAC 718 ITGA4 11222 CGGGCTGCATGCGTGAGCAGG 840 ITGA4 11252 CGGCAGGCGGTTTAGGCTGTGGCTG 885 ITGA4 11278 CCGATTCGGATTGCTCCAGCTGG 962 ITGA4 11289 CGCACCCACTCAGTTGCCACGGG 1008 ITGA4 11327 CGGAGACCCACAACGCAACACACC 1099 APP 7607 CGCGACCCTGCGCGGGGCACCG 1 APP 7741 GTGCGAGTGGGATCCGCCGCG 34 APP 7875 CGCGCCGCCACCGCCGCCGTCTCCCGG 68 APP 8009 CGCGCACGCTCCTCCGCGTGCTCTCG 101 APP 8143 CCGAGGAAACTGACGGAGCCCGAGCGCGG 137 APP 8145 CGAGTCAGCTGATCCGGCCCACCCCG 186 APP 8310 CGAGAGAGACCCCTAGCGGCGCCG 221 APP 8475 CGCCCGCTCGCGCCGGGAGGGGCCCTCG 256 APP 8640 CGCGCCCACAGGTGCACGCGCCCTTGGCG 289 APP 8805 GGCCGACGGCCCACCTGGGCTTCG 351 APP 8825 CGCTGAGGCTCTAGAAAAGTCGAGAG 446 APP 8843 CTCGTCCCCGTGAGCTTGAATCATCCGACCC 480 APP 8912 AGGCGTTTCTGGAAGAGAATGAGAACG 604 APP 8927 CGTCAAAAGCAGGCACGAGCAACCTG 701 APP 8928 GAACGAACCAAAGGAGCAAGGCG 742 APP 8929 CGCTGACAAGGGTGCCTAGGCCCGG 1318 APP 8948 CGCAATTCCGTATTTGTTCCGG 1738 APP 8969 GTACGTTGGCAGACGCAGTGACG 4923 CMYC 7551 CGATGAGGGTATTAACTCTGGC 335580 CMYC CM12 7552 CGGGGGTCCTCAGCCGTCCAGACC 518 CMYC CM13 7602 CGCTTATGGGGAGGGTGGGGAGGG 634 CMYC CM14 7603 CGGTGGGCGGAGATTAGCGAGAGA 559 CMYC 7606 GGCGCTTATGGGGAGGGTGGGGAGGG 632 CMYC 13684 CCTGGCACGTGTCCCTGGTCAAG 3482 CMYC 13703 CACGTGCGGCCTGTCAAGAGATGA 5926 FGFR1 13484 CGAGCCAGGCAGGGCCCCTCGCAAGTG 1850 FGFR1 13522 GACGGATATGAGTCCAGAAGTTGCG 1472 FGFR1 13535 TAGCTGCGTGCAGTGGCGCGCGCCTGT 4910 FGFR1 13561 CCGCCTCGCCAGCTCCCGAGCGCGAGTT 10239 FGFR1 13655 CGCCTCCTCCCAGGTGTGGGCTGGCTGCAGACCG 3067 CD68 13681 CGAGAACATGGCTTTCCAGCGTCTG 520 ALK 11471 CGCCGGAGGAGGCCGTTTACACTGC 3 ALK 11530 CGTGCGCGCAAGTCTCTTGCTTTCC 132 ALK 11555 CGCTCTCCGCGCCGAGTGCCGCGCC 269 ALK 11621 CGCCTTTTGCGTTCCTTTTGGCTCC 482 ALK 11681 CGCAGGCACTGGAGCGGCCCCGGCG 701 ALK 11794 CGACCCTCCGAACAGAGGCGGCGGG 851 ALK 11825 CGCGCTGCTGCCCGACCCACGCAGT 1022 ALK 11901 CGGGTCCGACTTCGGAAAAACAGGT 1313 ALK 11923 CGGCCTGTCGGGTAGCACAGGAGTT 2022 MSI2 11989 CGGTGACGTCACGCACCCCCGTGCG 360 MSI2 12058 CGGATACAATTACCCATATTGT 1535 MSI2 12059 GACTCAGTTGCTAACAACCATGAGCG 10624 MSI2 12060 CAGTTGCTAACAACCATGAGCG 10628 MSI2 12061 CATGAAAATTTCACCAAGTATAAATTAC 10909 MSI2 12062 CACCAAGTATAAATTACAGGTCT 10920 JAK2 12063 CGCACCAGTTTGTCCACGTCCAGTG 1663 JAK2 12098 GCCGTCACTGCCGACATAAGCACAGAC 1811 CCND1 12098 CGCTGCTACTGCGCCGACAGCCCTC 133 CCND1 12242 CGGCAGAATGGGCGCATTTCCAAGA 612 CCND1 12287 ACGCCACGAGGGCACCCACGGGCGGA 637 CCND1 12332 CGGTGACCGCGGCCTGGGCGGATGG 2755 CCND1 12388 CGGGACTCAGCGCGGCTGCGCGCCG 2907 BL9 13682 TGTCCACCTGAACACCTAGTCC 2388

TABLE 4 Additional DNAi Sequences Used in Supporting Data (disclosed in Pat. No.: 7,807,647) Location relative to 5′ upstream region Target Design ID Sequence ID Sequence from gene start site KRAS KR1 51 CCCGGAGCGGGACCGGACCGCGG 5923 KRAS KR2 52 GCCGGACCCACGCGGCGGCCCGCC 5856 BCL2 BL2 13724 CACGCACGCGCATCCCCGCCCGTG 2388 BCL2 BL3 13725 ACCGGCGCTCGGCGCGCGGA Mismatched BCL2 BL4 13726 GACGCGCCGGGCCGGGCGGA Mismatched BCL2 BL7 13727 GGCGCGCGGGGCCGGGCCGGG — CMYC CM7 13728 GGGCGCCTCGCTAAGGCTGGGGAAAGGGCCGCGC  969

TABLE 5 DNAi Sequences Used in Supporting Data as Negative Controls Location relative to 5′ upstream Target Design ID SEQ ID NO: Sequence region from gene start site Survivin SU3 105 GACATCGCTGTCCCGGCGAGTACATCGTT 665 KRAS KR0525 1516 AGTCTCCCCTTCCCGGAGACT 10265 

1. An oligonucleotide that hybridizes to a non-coding region in or upstream of a promoter for a target gene, wherein the oligonucleotide comprises: a length of 20-34 bases; at least one CG pair; at least 40% C and G content; no more than five consecutive bases of the same nucleotide; and at least one secondary structure for said oligonucleotide.
 2. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a C and G content of at least 50%.
 3. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a C and G content from about 50 to 80%.
 4. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least two CG pairs.
 5. The oligonucleotide of claim 1, wherein said oligonucleotide hybridizes within a CG region, CpG island region, nuclease hypersensitive site, or CIS regulatory region.
 6. The oligonucleotide of claim 1, wherein said non-coding region is located within a CG region, CpG island, nuclease hypersensitive site, or CIS regulatory region.
 7. The oligonucleotide of claim 1, wherein said oligonucleotide is a reverse and full complement of a sense strand of said non-coding region of the target gene.
 8. The oligonucleotide of claim 1, wherein said oligonucleotide is unique to the nucleotide sequence of the non-coding region.
 9. The oligonucleotide of claim 1, wherein the nucleotide sequence of the non-coding region is not duplicated in a genome comprising the target gene.
 10. The oligonucleotide of claim 1, wherein the nucleotide sequence of the non-coding region comprises less than 80% homology to other nucleotide sequences in a genome with a target gene.
 11. The oligonucleotide of claim 1, wherein the nucleotide sequence of the non-coding region comprises less than 50% homology to other nucleotide sequences in a genome with a target gene.
 12. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least four bases in a linear section of the secondary structure.
 13. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least five bases in a linear section of the secondary structure.
 14. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one CG pair within the first 40% of the bases of said oligonucleotide.
 15. The oligonucleotide of claim 1, wherein said oligonucleotide comprises at least one CG pair within the first 50% of the bases of said oligonucleotide.
 16. The oligonucleotide of claim 1, wherein said oligonucleotide further comprises at least one CG pair that is prior to or in the nonlinear section of the secondary structure.
 17. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a linear section before a secondary structure, no oligonucleotides that extend beyond the secondary structure, and at least one CG pair within the linear section or the secondary structure.
 18. The oligonucleotide of claim 1, wherein said oligonucleotide has a linear section before a secondary structure and no oligonucleotides that extend beyond the secondary structure
 19. The oligonucleotide of claim 1, wherein said oligonucleotide does not comprise a single G or T base after the nonlinear section of the secondary structure.
 20. The oligonucleotide of claim 1, wherein said secondary structure comprises at least one hairpin loop.
 21. The oligonucleotide of claim 1, wherein said secondary structure comprises at least two hairpin loops.
 22. The oligonucleotide of claim 19 or 20, wherein said secondary structure comprises at least three nucleotide bridges in the nonlinear section of the secondary structure.
 23. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a theoretical ΔG between −0.1 to −7.
 24. The oligonucleotide of claim 23, wherein said theoretical ΔG is between −1 to −5.
 25. The oligonucleotide of claim 1, wherein said oligonucleotide comprises a theoretical ΔTm between 30-70 degrees Celsius.
 26. The oligonucleotide of claim 1, wherein said oligonucleotide begins at the 5′ end with the bases selected from CG, CGG, CGC, CGT, CGA, GCG, CCC, CCG, GTC, TCC, TCG, ACG, CAC, CAG, GAG, AGA, GAC, GAA, AGC, or GCC.
 27. The oligonucleotide of claim 1, wherein said oligonucleotide ends at the 3′ end with the bases selected from CG, GCG, GGC, CGG, GCC, CGC, CCG, ACG, TCG, GGG, TGC, CCC, GTG, or CTC.
 28. The oligonucleotide of claim 1, wherein said non-coding region is located less than 7000 bases upstream of the coding region of the target gene.
 29. The oligonucleotide of claim 1, wherein said non-coding region is located less than 5000 bases upstream of the coding region of the target gene.
 30. The oligonucleotide of claim 1, wherein said non-coding region is located less than 3000 bases upstream of the coding region of the target gene.
 31. The oligonucleotide of claim 1, wherein said non-coding region is located less than 1000 bases upstream of the coding region of the target gene.
 32. The oligonucleotide of claim 1, wherein said non-coding region is located less than 500 bases up- or downstream of a transcription factor binding site or translocation site of target gene.
 33. The oligonucleotide of claim 1, wherein said non-coding region is located less than 100 bases up- or downstream of a transcription factor binding site or translocation site of target gene.
 34. The oligonucleotide of claim 1, wherein said oligonucleotide does not comprise a CpG Coley motif.
 35. The oligonucleotide of claim 1, wherein said oligonucleotide does not form a triplex structure.
 36. The oligonucleotide of claim 1, wherein said oligonucleotide does not form a G-quadruplex structure.
 37. The oligonucleotide of claim 1, wherein said oligonucleotide is a single stranded DNA.
 38. The oligonucleotide of claim 1, wherein said oligonucleotide hybridizes to an Sp1 motif or transcription factor binding site.
 39. The oligonucleotide of claim 1, wherein said target gene is selected from Survivin, Beclin-1, STAT3, HIF1A, IL-8, KRAS, MTTP, ApoC III, ApoB, IL-17, MMP2, FAP, P-selectin, IL-6, IL-23, AKT, CRAF, Beta-catenin, PCSK9, MEK1, MEK2, CD4, WNT1, Clusterin, NRAS, EZH2, HDAC1, PD-1, TNFα, MIF1, TTR, HBV, HAMP, ERBB2, PARP1, ITGA4, APP, FGFR1, CD68, ALK, MSI2, JAK2, CCND1, or selected from Table
 2. 40. The oligonucleotide of claim 1, wherein said oligonucleotide is selected from the group consisting of any of the sequences disclosed in Table
 3. 41. The oligonucleotides of claim 1, wherein said oligonucleotide hybridizes to a hot zone of a target gene.
 42. The oligonucleotide of claim 1, wherein at least one of the cytosine bases in said oligonucleotide is 5-methylcytosine.
 43. The oligonucleotide of claim 1, wherein at least one of the cytosine bases in said CG pair is 5-methylcytosine.
 44. The oligonucleotide of claim 1, wherein all of said cytosine bases in said oligonucleotide are 5-methylcytosine.
 45. The oligonucleotide of claim 1, wherein said hybridization of said oligonucleotide to the non-coding region modulates the target gene.
 46. The oligonucleotide of claim 1, wherein said hybridization of said oligonucleotide to the non-coding region of the target gene modulates expression or transcription of said target gene.
 47. The oligonucleotide of claim 1, wherein said hybridization of said oligonucleotide to the non-coding region of the target gene modulates a cell signaling pathway.
 48. The oligonucleotide of claim 1, wherein said hybridization of said oligonucleotide to the non-coding region of said target gene produces phenotypic changes in a mammal.
 49. The oligonucleotide of claim 1, wherein said hybridization of said oligonucleotide to the non-coding region of said target gene influences a non-gene target due to a chromosomal rearrangement.
 50. The oligonucleotide of claim 1, wherein said target gene is on a chromosome of a cell, and wherein said hybridization of said oligonucleotide to said non-coding region reduces proliferation of said cell.
 51. The oligonucleotide of claim 1, wherein said target gene is an oncogene.
 52. A composition comprising an oligonucleotide according to any one of claims 1-51 and a pharmaceutically acceptable carrier.
 53. The composition of claim 52, wherein the pharmaceutically acceptable carrier is a liposome.
 54. The composition of claim 53, wherein the liposome is an amphoteric liposome.
 55. The composition of claim 53, wherein the liposome comprises a neutral lipid.
 56. The composition of claim 53, wherein the liposome comprises a mixture of neutral lipids and lipids with amphoteric properties, wherein the mixture of lipid components comprises anionic and cationic properties and at least one such component is pH responsive.
 57. The composition according to any one of claims 52-56, wherein the composition further comprises an additional therapeutic agent.
 58. The composition of claim 57, wherein the additional therapeutic agent is a second oligonucleotide, chemotherapeutic agent, immunotherapeutic agent, or radiotherapy.
 59. The composition of claim 52, wherein said composition has two (2) therapeutic agents.
 60. The composition of claim 59, wherein one therapeutic agent treats a cancer disease and the other therapeutic agent treats a non-cancer disease.
 61. A method of inhibiting protein expressing in a cell with a target gene comprising introducing into said cell an oligonucleotide according to any one of claims 1-51 or composition according to any one of claims 52-60.
 62. A method of mediating target-specific RNA in a mammalian cell in vitro, comprising contacting said mammalian cell in vitro with an oligonucleotide according to any one of claims 1-51 or composition according to any one of claims 52-60.
 63. A method of mediating protein down regulation in a mammalian cell in vitro, comprising contacting said mammalian cell in vitro with an oligonucleotide according to any one of claims 1-51 or composition according to any one of claims 52-60.
 64. A method of treating a patient having a disease characterized by the presence or undesired production of a protein implicated in said disease, comprising administering to said patient a pharmaceutically effective amount of an oligonucleotide according to any one of claims 1-51 or composition according to any one of claims 52-60.
 65. A method of treating a patient having a disease characterized by the presence or undesired production of a protein implicated in said disease, comprising administering to said patient a pharmaceutically effective amount between 1 mg/m² and 500 mg/m² of an oligonucleotide according to any one of claims 1-51 or composition according to any one of claims 52-60.
 66. A method of treating a mammal having a disease characterized by the presence or undesired production of a protein implicated in disease, comprising administering to said mammal a pharmaceutically effective amount of an oligonucleotide according to any one of claims 1-51 or composition according to the description and the compositions in any of claims 52-60.
 67. A method of treating a plant having a disease characterized by the presence or undesired production of a protein implicated in disease, comprising introducing to said plant an effective amount of an oligonucleotide according to any one of claims 1-51 or composition according to the description and the compositions in any of claims 52-60.
 68. A method of administration of a therapeutic disclosed herein and a oligonucleotide according to any one of claims 1-51 or a composition according to any one of claim 52-60, wherein said administration is through a route selected from oral, vapor, inhalation, dermal, subdermal, subcutaneous, parental, parenterally, ear, nose, nasally, bucally, eye, otic, ophthalmically, rectal, vaginal, suppository or implant, implanted reservoir, dermal, dermal skin patch, injection, or sub-lingual.
 69. A method or kit for a diagnosis and treatment of a disease comprising the steps of administering to a patient a pharmaceutically effective amount of an oligonucleotides accordingly to any one of claims 1-51 or a composition according to any one of claims 52-60, wherein the patient is characterized by the presence of, or undesired production of, a protein implicated in said disease, and the method further comprising evaluating said patient for the presence of, or undesired production of said protein.
 70. An single stranded DNA oligonucleotide that hybridizes to coding or non-coding region of a target gene, wherein the oligonucleotide comprises: a length of 12-50 bases; at least 30% C and G content; and no more than seven consecutive bases of the same nucleotide.
 71. The oligonucleotide of claim 70, wherein the nucleotide sequence of the non-coding region comprises less than 80% homology to other nucleotide sequences in a genome with a target gene.
 72. The oligonucleotide of claim 70, wherein said oligonucleotide comprises at least one CG pair within the first 40% of the bases of said oligonucleotide.
 73. The oligonucleotide of claim 70 further comprising a secondary structure.
 74. The oligonucleotide of claim 70, wherein said oligonucleotide comprises a theoretical ΔG between −0.1 to −7.
 75. The oligonucleotide of claim 70, wherein said oligonucleotide comprises a theoretical ΔTm between 30-70 degrees Celsius.
 76. The oligonucleotide of claim 70, wherein said non-coding region is located less than 7000 bases upstream of the coding region of the target gene.
 77. The oligonucleotide of claim 70, wherein said non-coding region is located less than 500 bases up- or downstream of a transcription factor binding site or translocation site of target gene.
 78. The oligonucleotide of claim 70, wherein said non-coding region is located with a CG region, nuclease hypersensitive site, or CpG island of the genome comprising the target gene.
 79. The oligonucleotide of claim 70, further comprises at least one CG pair and optionally at least one of the cytosine bases in said CG pair is 5-methylcytosine.
 80. The oligonucleotide of claim 70, wherein said target gene is on a chromosome of a cell, and wherein said hybridization of said oligonucleotide reduces proliferation of said cell.
 81. A composition comprising an oligonucleotide according to any one of claims 70-80 and a pharmaceutically acceptable carrier.
 82. The composition of claim 81, wherein the pharmaceutically acceptable carrier is a liposome.
 83. The composition according to any one of claim 81 or 82 wherein the composition further comprises an additional therapeutic agent.
 84. A method of inhibiting or silencing gene transcription in a cell with a target gene comprising introducing into said cell an oligonucleotide according to any one of claims 70-80 or composition according to any one of claims 81-83.
 85. A method of mediating target-specific RNA in a mammalian cell in vitro, comprising contacting said mammalian cell in vitro with an oligonucleotide according to any one of claims 70-80 or composition according to any one of claims 81-83. 